Figure S1. General experimental procedure
98
Figure S1. General experimental procedure RT H2B H4 H2A H3 Intenti sity Equal mixture of L&H histones Step 1. Propionylatio n Step 2. Trypsin digestion Step 3. Propionylatio n NanoLC-MS/MS analysis Intenti sity M/Z Nuclei prep/Aci d Extracti on WT ΔEZL2 ΔTXR1 N15 WT RP-18 HPLC separation
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Figure S1. General experimental procedure. WT. Nuclei prep/Acid Extraction. H2B. Intentisity. Step 1. Propionylation Step 2. Trypsin digestion Step 3. Propionylation. Intentisity. ΔEZL2. H4. H2A. H3. ΔTXR1. RT. M/Z. Equal mixture of L&H histones. N15 WT. - PowerPoint PPT Presentation
Transcript of Figure S1. General experimental procedure
Figure S1. General experimental procedure
RT
H2B
H4H2A
H3Inte
ntis
ity
Equal mixture of L&H histones
Step 1. Propionylation Step 2. Trypsin digestionStep 3. Propionylation
NanoLC-MS/MS analysis
Inte
ntis
ity
M/Z
Nuclei prep/Acid Extraction
WT
ΔEZL2
ΔTXR1
N15 WT RP-18 HPLC separation
a. H2A variants
H2A.X/H2A.1:K5Ac, K8Ac, K10Ac, K12Ac, S1/T2/T3Phos*, R33Me1, R36Me2, K39Form, K42Me1, R92Me1H2A.Z:K4Ac,K7Ac,K10Ac,K13Ac,K16Ac,K21Ac,K140Ac
H2A.X/H2A.1:K5Ac, K8Ac, K10Ac, K12Ac, K39Form, R92Me1H2A.Z:K4Ac,K7Ac,K10Ac,K13Ac,K16Ac,K21Ac,K140Ac
*Localization is difficult due to incomplete fragment ion information
Figure S2. Histone variants, sequence coverage, and their PTMs
# of PTMs quantified (13 PTMs):
# of PTMs identified (17 PTMs):
H2A --------STTGKGGKAKG-KTASSKQVSRSARAGLQFPVGRISRFLKNGRYS-ERIGTG 50H2A.X --------STTGKGGKAKG-KTASSKQVSRSARAGLQFPVGRISRFLKHGRYS-ERVGTG 50 H2A.Z AGGKGGKGGKGGKGGKVGGAKNKKTPQ-SRSYKAGLQFPVGRIHRFLKGRVSAKNRVGAT 59
H2A APVYLAAVLEYLAAEVLELAGNAAKDNKKTRIVPRHILLAIRNDEELNKLMANTTIADGG 110H2A.X APVYLAAVLEYLAAEVLELAGNAAKDNKKTRIVPRHILLAIRNDEELNKLMANTTIADGG 110H2A.Z AAVYAAAILEYLTAEVLELAGNASKDFKVRRITPRHLLLAIRGDEEL-DILIKATIAGGG 118 H2A VLPNINPMLLPSKTKKS-----TEPEH 132H2A.X VLPNINPMLLPSKSKKTESRGQASQDL 137H2A.Z VIPHIHKALLGKHSTKNRSSAKTAEPR 145
Sequence coverage: 60%
b. H2B variants
H2B1 APKKAPAAAAEKKVKKAPTTEKKNKKKRSETFAIYIFKVLKQVHPDVGISKKAMNIMNSF 60H2B2 APKKAPAATTEKKVKKAPTTEKKNKKKRSETFAIYIFKVLKQVHPDVGISKKAMNIMNSF 60 H2B1 INDSFERIALESSKLVRFNKRRTLSSREVQTAVKLLLPGELARHAISEGTKAVTKFSSST 120H2B2 INDSFERIALESSKLVRFNKRRTLSSREVQTAVKLLLPGELARHAISEGTKAVTKFSSSS 120 H2B1 N 121H2B2 N 121
Sequence coverage: 68%
H2B-1: A1Me1, K3Me1/Me2/Me3, K4Ac, K115Ac, (S117-T120)Phos*
H2B-2: A1Me1, K3Me1/Me2/Me3, K4Ac
H2B-1: A1Me1, K3Me1/Me2/Me3, K4Ac, K115Ac, (S117-T120)Phos* H2B-2: K3Me2/Me3, K4Ac
# of PTMs quantified (10 PTMs):
*Localization is difficult due to incomplete fragment ion information
# of PTMs identified (12 PTMs):
ARTKQTARKSTGAKAPRKQLASKAARKSAPATGGIKKPHRFRPGTVALREIRKYQKSTDL 60ARTKQTARKSTGVKAPRKQLATKAARKSAPVSGGVKKPHKFRPGTVALREIRKYQKTTDL 60ARTKQTARKSTSIKAPRKQLAAKAARKSAPISGGIKKPHKFRPGTVALREIRKYQKTTDL 60
LIRKLPFQRLVRDIAHEFKAELRFQSSAVLALQEAAEAYLVGLFEDTNLCAIHARRVTIM 120LIRKLPFQRLVRDIAMEMKSDIRFQSQAILALQEAAEAYLVGLFEDTNLCAIHARRVTIM 120LIRKLPFQRLVRDIAMEMKSDIRFQSQAILALQEAAEAYLVGLFEDTNLCAIHARRVTIM 120
TKDMQLARRIRGERF 135TKDLHLARRIRGERF 135TKDLHLARRIRGERF 135
H3 H3.3 H3.4 H3 H3.3 H3.4 H3 H3.3 H3.4
c. H3 variants
Sequence coverage: 76%
H3: K4Me1/Me2/Me3, K9Ac, K14Ac, K14Me1, K18Ac, K23Ac, K23Me1/Me2/Me3, K23Form, K27Me1/Me2/Me3, K27Ac, K36Me1/Me2, R40Me1, R49Me1, K56Me1, K79Me1, R83Me1, R128Me1
H3: K4Me1/Me3, K9Ac, K14Ac, K18Ac, K23Ac, K23Me1/Me2/Me3, K23Form, K27Me1/Me2/Me3, K27Ac, K36Me1, K56Me1, R83Me1, R128Me1
# of PTMs identified (32 PTMs):
H3.3: K9Ac, K14Ac, K14Form, K18Ac, K23Ac, K23Form, K56Ac, K79Me1
# of PTMs quantified (25 PTMs):
H3.3: K9Ac, K14Ac, K14Form, K18Ac, K23Ac, K23Form, K56Ac
d. H4
AGGKGGKGMGKVGAKRHSRKSNKASIEGITKPAIRRLARRGGVKRISSFIYDDSRQVLKS 60
FLENVVRDAVTYTEHARRKTVTAMDVVYALKRQGRTLYGFGG 102
Sequence coverage: 97%
H4: K4Ac, K7Ac, K7Ub, K11Ac, K15Ac, R35Me1, R55Me1, R67Me1, R77Me1, K91Me1, R92Me1
H4: K4Ac, K7Ac, K11Ac, K15Ac, R55Me1, R77Me1, K91Me1# of PTMs quantified (7 PTMs):
# of PTMs identified (11 PTMs):
WT1_H2A_propionylated_1 #2586-7074 RT: 40.14-75.94 AV: 9 NL: 1.55E2T: Average spectrum MS2 572.81 (2586-7074)
200 300 400 500 600 700 800 900 1000 1100m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Re
lative
Ab
un
da
nce
554.91
871.50
942.37274.26
370.17
499.25563.85203.22
299.23 481.33
853.12 924.36630.19237.34 959.55687.24 756.56 1029.42
M/Z: 572.8141, Charge: 2+, Mass Error: 0ppm, Ion Score: 23
H2A.X-K5Ac1STTGKGGKAK10
y1
y2
y3y4
y5
b6 b7
b8
b9
Figure S3. All PTMs identified by tandem MS
[M-2H2O+2H]2+
M/Z: 1172.628, Charge: 2+, Mass Error: 1ppm, Ion Score: 54
H2A.X-K5AcK8AcK10AcK12Ac1STTGKGGKAKGKTASSKQVSR21
WT1_H2A_propionylated_1 #2835-2853 RT: 42.42-42.58 AV: 3 NL: 1.59E2T: Average spectrum MS2 1173.13 (2835-2853)
400 600 800 1000 1200 1400 1600 1800 2000m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100R
ela
tive
Ab
un
da
nce
1154.61
847.63
1479.571246.83
1098.421019.73
1325.49
918.66
760.63 1430.96
1837.75
1501.82 1786.03
489.31 673.58
1729.10361.32 1984.511566.55655.38
412.37
y3
y4 y5
y6
y7
y8
y9
y11
y12 y13
b13
b14-H2O
b15-H2O
b16-H2Ob16
b17-H2O
b18
[M-2H2O+2H]2+
M/Z: 786.7592, Charge: 3+, Mass Error: 0ppm, Ion Score: 34
H2A.X-K5AcK8AcK10Ac1STTGKGGKAKGKTASSKQVSR21
WT1_H2A_propionylated_2 #2846-9195 RT: 44.17-96.54 AV: 26 NL: 2.58E2T: Average spectrum MS2 787.09 (2846-9195)
400 600 800 1000 1200 1400 1600 1800 2000m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100R
ela
tive
Ab
un
da
nce
768.68
1049.08
847.54
1092.56
489.40 1310.67
670.35
900.51262.20
1019.58361.381112.42
1260.72624.03 1493.491211.08
426.71 1598.311851.72
y2
y3
y4
y5
y6
y7
y8
y8-H2O
y9
y10
y19-NH3
b8
b7-NH3
b12
b20
[M-2H2O+2H]2+
M/Z: 1179.6361, Charge: 2+, Mass Error: 1ppm, Ion Score: 69
H2A.X-K5AcK8AcK12Ac1STTGKGGKAKGKTASSKQVSR21
WT1_H2A_propionylated_2 #2863-2879 RT: 44.30-44.42 AV: 3 NL: 2.10E2T: Average spectrum MS2 1180.14 (2863-2879)
400 600 800 1000 1200 1400 1600 1800 2000m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100R
ela
tive
Ab
un
da
nce
1161.66
1246.80847.60 1339.49
1493.51
1019.57
1112.52
1440.56918.67760.56 1851.67
1979.911598.46673.57
1667.89489.43361.61 573.20
y3y4
y5
y6
y7
y8
b14-H2O
y9
y11
b12
b15y12
y13 b16-H2O
b17-H2O b18-H2O
b18
[M-2H2O+2H]2+
M/Z: 1186.6435, Charge: 2+, Mass Error: 1ppm, Ion Score: 101
H2A.X-K5AcK8Ac1STTGKGGKAKGKTASSKQVSR21
WT2_H2A_propionylated_2 #2645 RT: 44.35 AV: 1 NL: 3.58E2T: ITMS + c NSI d Full ms2 [email protected] [315.00-2000.00]
400 600 800 1000 1200 1400 1600 1800 2000m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Re
lativ
e A
bu
nd
an
ce
1168.65
1019.60
1353.32
1122.25847.55 1260.66811.49
1507.781594.72 1847.70918.56617.67 760.63 1700.03489.29
1883.82
y4y5
y6
y7
y8
b14-H2O
y9
y11
b10
b12
y12y13
b15-H2O
b13y16
b15b14
[M-2H2O+2H]2+
M/Z: 1212.6277, Charge: 2+, Mass Error: 1ppm, Ion Score: 43
H2A.X-STT(1-3)Phos1STTGKGGKAKGKTASSKQVSR21
WT1_H2A_propionylated_1 #3157-4004 RT: 45.02-51.86 AV: 6 NL: 1.30E4T: Average spectrum MS2 1213.09 (3157-4004)
400 600 800 1000 1200 1400 1600 1800 2000m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
1190.92
1137.781045.64 1281.70 1565.831493.74 1653.74933.61 1741.91 1936.09849.35742.60668.47583.42479.32 y6 y7y9
y11b5 b12
[M-H2O+2H]2+
b6
b9
b10-NH3
b13 b14 b15 b16
b17
M/Z: 893.4959, Charge: 2+, Mass Error: 2ppm, Ion Score: 72
H2A.X-K10AcK12Ac6GGKAKGKTASSKQVSR21
TRR1_1_H2A_propionylated_1 #2471-2486 RT: 41.76-41.89 AV: 3 NL: 2.22E2T: Average spectrum MS2 893.50 (2471-2486)
400 600 800 1000 1200 1400 1600 1800m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100R
ela
tive
Ab
un
da
nce
1246.69
1019.62
847.62
875.64
918.64
1524.79
760.60
1430.83749.47
489.44540.40
716.261279.60
361.41 673.561189.75 1611.81
262.281113.68
597.47472.36373.34
1685.23
y4
y5
y6
y7
y8
b12-H2O
y9
y11
b5
b12y10
b15-H2O
b13b15
b14
y2
y3
b6
b7
b10b11
b14-H2O
M/Z: 900.504, Charge: 2+, Mass Error: 2ppm, Ion Score: 79
H2A.X-K10Ac6GGKAKGKTASSKQVSR21
WT1_H2A_propionylated_1 #3062-4827 RT: 44.25-58.35 AV: 5 NL: 7.47E1T: Average spectrum MS2 900.50 (3062-4827)
400 600 800 1000 1200 1400 1600 1800m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100R
ela
tive
Ab
un
da
nce
882.54
847.69
1019.73
530.42
1538.62
489.391260.84
643.52 760.64 935.29
1625.94600.45 1444.68382.33 1189.76
1293.73348.30 1165.77
y4
y5
y6
y7
y9
y11
b12
y10b13
b15
b14
y2 y3b6
b7
b10
b14-H2O
y12
[M-2H2O+2H]2+
M/Z: 912.4688, Charge: 2+, Mass Error: 2ppm, Ion Score: 58
H2A.X-K12Ac6GGKAKGKTASSKQVSR21
WT2_H2A_propionylated_2 #2501-4293 RT: 43.14-58.09 AV: 6 NL: 8.76E1T: Average spectrum MS2 912.47 (2501-4293)
400 600 800 1000 1200 1400 1600 1800m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100R
ela
tive
Ab
un
da
nce
903.39
859.53
1033.57
1277.78
931.44
497.35 773.25
1557.641645.70
683.32 1463.761053.45 1327.59
1667.08561.36267.39 1209.75601.72479.78369.24
1787.16
y4
y5
y6
y7
y9
y11
b11y10
b13
b15b14
b6b10
y13
b5
b6-NH3
b12
b12-NH3
b14-NH3
[M-H2O+2H]2+
M/Z: 514.2709, Charge: 2+, Mass Error: 0ppm, Ion Score: 30
H2A.X-R33Me125AGLQFPVGR33
WT1_H2A_propionylated_1 #5299 RT: 62.05 AV: 1 NL: 2.35E3T: ITMS + c NSI d Full ms2 [email protected] [130.00-1040.00]
200 300 400 500 600 700 800 900 1000m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Re
lativ
e A
bu
nd
an
ce
504.70
597.47
727.53
449.46
579.36
301.14
251.19
431.11777.19356.46
851.53421.35 709.58651.09193.16 881.60520.20 1003.70
y4
y5
y6
b7b4
b6
b3
b5
y2
y1
y3y8
[M-H2O+2H]2+
TRR1_1_H2A_propionylated_1 #6206 RT: 71.16 AV: 1 NL: 2.67E2T: ITMS + c NSI d Full ms2 [email protected] [160.00-1270.00]
200 300 400 500 600 700 800 900 1000 1100 1200m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100R
ela
tive
Ab
un
da
nce
618.54542.91
369.27
813.48
666.48
1084.64254.02499.28317.16 1022.32423.08
232.17 715.94886.20440.63 780.67 994.60
1231.35
H2A.X-R36Me234ISRFLKHGR42
M/Z: 627.3735, Charge: 2+, Mass Error: -6ppm, Ion Score: 43
y4
y5
y6
b7
b6
b5
y2
y3
y8
y7
y7,2+
y8,2+
b3-H2O
b8
[M-H2O+2H]2+
H2A.X-R42Me137FLKHGR42
M/Z: 448.2434, Charge: 2+, Mass Error: 0ppm, Ion Score: 39
WT1_H2A_propionylated_1 #3115-4623 RT: 44.65-56.90 AV: 4 NL: 6.68E3T: Average spectrum MS2 447.75 (3115-4623)
200 300 400 500 600 700 800 900m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Re
lativ
e A
bu
nd
an
ce
391.25
346.32
577.47691.53
251.16439.33
289.17
645.28301.18205.07
141.10 505.22703.31617.35487.41 833.58798.83 875.09
y4y5
b3
y2
y3
y5,2+
b1
[M-H2O+2H]2+
b2
b4
b5
H2A.X-K39Form37FLKHGR42
M/Z: 421.2374, Charge: 2+, Mass Error: 0ppm, Ion Score: 36
WT1_H2A_propionylated_1 #3671 RT: 49.26 AV: 1 NL: 1.21E2T: ITMS + c NSI d Full ms2 [email protected] [105.00-855.00]
200 300 400 500 600 700 800m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Re
lativ
e A
bu
nd
an
ce
319.81
525.51
403.36
638.55
369.43
232.29412.37
302.30175.98 296.13
140.00 246.10 685.57199.11553.73 621.34
422.17 508.56
y4
y5
y2
y3
y5,2+
b1
[M-2H2O+2H]2+
b2 b5
y1
b4-NH3b4,2+b5-NH3
H2A.X-R92Me186HILLAIR92
M/Z: 917.5589, Charge: 1+, Mass Error: 2ppm, Ion Score: 53
WT1_H2A_propionylated_1 #6430-6451 RT: 70.77-70.94 AV: 3 NL: 8.03E2T: Average spectrum MS2 917.56 (6430-6451)
250 300 350 400 450 500 550 600 650 700 750 800 850 900m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100R
ela
tive
Ab
un
da
nce
611.33
539.26
425.23
311.15
899.56
725.43
873.67589.51
697.45493.41855.72439.19411.33
553.34361.35289.26 733.34625.30 844.23 917.91
y4
y5y3 y6
b3
b2
b4
b5
b6
[M-H2O+2H]2+
H2A.Z-K7AcK10Ac1AGGKGGKGGKGGK13
M/Z: 571.8189, Charge: 2+, Mass Error: 3ppm, Ion Score: 58
WT_2_G_2 #790 RT: 23.55 AV: 1 NL: 1.36E3T: ITMS + c NSI d Full ms2 [email protected] [145.00-1155.00]
200 300 400 500 600 700 800 900 1000 1100m/z
0
10
20
30
40
50
60
70
80
90
100
Rel
ativ
e A
bund
ance
829.07
562.55
507.55
479.20
772.18598.02
882.061014.05260.92 938.93186.07 370.79 715.12451.94
313.99 622.15252.32 y4
y5
y3y7
b3
b4
b5
b7
y8
y9
y11
y10,2+
y11,2+
b9
b10b11
b12
H2A.Z-K4AcK7AcK10Ac1AGGKGGKGGKGGK13
M/Z: 592.8227, Charge: 2+, Mass Error: 0ppm, Ion Score: 68
WT_2_G_2 #957-965 RT: 27.79-27.92 AV: 3 NL: 4.74E3T: Average spectrum MS2 592.82 (957-965)
200 300 400 500 600 700 800 900 1000 1100 1200m/z
0
10
20
30
40
50
60
70
80
90
100
Rel
ativ
e A
bund
ance
829.08
583.53
528.50772.09
622.04500.04 924.04
260.93 640.06715.08355.78
679.09412.87284.91 1038.06980.97202.86 1055.96341.95 888.61 1144.49
y4 y5
y3
y7b4
b5
b7
y8
y9
y11
y10,2+
y11,2+
b9
b10
b11 b12
y6
b6
b8
H2A.Z-K4AcK7AcK10AcK13Ac1AGGKGGKGGKGGKGGK16
M/Z: 734.897, Charge: 2+, Mass Error: 0ppm, Ion Score: 77
WT_1_G_1 #1437-1442 RT: 34.70-34.81 AV: 3 NL: 2.34E3T: Average spectrum MS2 734.90 (1437-1442)
200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400m/z
0
10
20
30
40
50
60
70
80
90
100
Rel
ativ
e A
bund
ance
829.01
1113.07
545.06
725.58
670.57622.07
906.03
1208.03924.00 1056.09
772.11
999.15488.11260.941264.97451.92355.90
1340.08y4
y5y3
b4
b5
b7 y8
y9
y11
y14,2+
b9
b10
b11 b12
y6
y10
y12
y14
b13
b14
b15
H2A.Z-K4AcK7AcK10AcK13AcK16Ac1AGGKGGKGGKGGKGGKVGGAK21
M/Z: 962.0239, Charge: 2+, Mass Error: 0ppm, Ion Score: 97
WT_2_G_3 #1620 RT: 44.33 AV: 1 NL: 1.56E3T: ITMS + c NSI d Full ms2 [email protected] [250.00-1940.00]
400 600 800 1000 1200 1400 1600 1800m/z
0
10
20
30
40
50
60
70
80
90
100
Rel
ativ
e A
bund
ance
1283.12
998.96 1591.02
715.06
897.58
1492.03
1208.01332.06431.09 640.03 784.21
1648.051453.051168.92
487.00 1776.061304.09
y4y5
b4b5
b7
y8
y9
y11
y14,2+
b11 b12
y7
y10
y12
y14
b13
b15
y13
y15
y17
y17,2+
y19,2+
b16
b17
b18
b19
b20
H2A.Z-K4AcK7AcK10AcK13AcK16AcK21Ac1AGGKGGKGGKGGKGGKVGGAKNK23
M/Z: 736.4026, Charge: 3+, Mass Error: 2ppm, Ion Score: 91
WT_2_G_2 #1682 RT: 44.66 AV: 1 NL: 3.51E3T: ITMS + c NSI d Full ms2 [email protected] [190.00-2000.00]
200 400 600 800 1000 1200 1400 1600 1800 2000m/z
0
10
20
30
40
50
60
70
80
90
100
Rel
ativ
e A
bund
ance
796.04
616.04
746.48715.02
973.611208.02
824.44604.42
1338.891055.07559.05284.95 355.84 1492.081112.09 1265.12 1590.91 1793.04y2
y5
b4 b7
y6
y11
b10b12
y7
y10b13
b15y13
b17,2+
y16,2+
b16b17
b18y3
y18,2+y19,2+
b6b14
b16,2+
b18,2+
H2A.Z-K7AcK10AcK13Ac5GGKGGKGGKGGK16
M/Z: 557.3039, Charge: 2+, Mass Error: 0ppm, Ion Score: 67
WT_1_G_1 #1123 RT: 28.44 AV: 1 NL: 2.31E3T: ITMS + c NSI d Full ms2 [email protected] [140.00-1125.00]
200 300 400 500 600 700 800 900 1000 1100m/z
0
10
20
30
40
50
60
70
80
90
100
Re
lativ
e A
bu
nd
an
ce
829.01
548.05
500.08284.95
853.02569.07 772.23260.85
715.01487.97
608.06341.91967.08398.97 909.78665.13 985.32207.88
y14,2+
y4
y5b4
b5
b7
y8
y9
y6
b11
b3
y7
y10,2+
y3
y10
b6
b9
b10
H2A.Z-K7AcK10AcK13AcK16Ac5GGKGGKGGKGGKVGGAK21
M/Z: 784.4312, Charge: 2+, Mass Error: 0ppm, Ion Score: 78
WT_2_G_2 #1450 RT: 39.52 AV: 1 NL: 3.62E3T: ITMS + c NSI d Full ms2 [email protected] [205.00-1580.00]
400 600 800 1000 1200 1400m/z
0
10
20
30
40
50
60
70
80
90
100
Re
lativ
e A
bu
nd
an
ce
999.13
715.17 1236.06835.03
1283.04332.00
1218.07568.94 727.13 853.01
1421.06642.02431.02 775.05 934.11 1118.96
284.95
1307.07398.87
455.14 1065.82 1439.03
y4
y5
b5
y8
y9
b13
b3
y7
y14,2+
y3
y10
b6b9
b12
y11
y12
y13
y14
y15,2+
b14
b15
b16
H2A.Z-K7AcK10AcK13AcK16AcK21Ac5GGKGGKGGKGGKVGGAKNK23
M/Z: 618.005, Charge: 3+, Mass Error: -2ppm, Ion Score: 61
WT_2_G_1 #1579 RT: 42.68 AV: 1 NL: 3.67E3T: ITMS + c NSI d Full ms2 [email protected] [160.00-1870.00]
200 400 600 800 1000 1200 1400 1600 1800m/z
0
10
20
30
40
50
60
70
80
90
100
Rel
ativ
e A
bund
ance
569.04
714.99
642.20
852.93984.09
784.14
1137.03261.01 431.00909.92
1438.071236.26
y5
b5 y8
b13b3
y7
y14,2+
y3
y10
b6
b9
b12
y12
y16,2+
y2
y7,2+ y10,2+
y13,2+
b10b14,2+
b17,2+
H2A.Z-K13AcK16Ac8GGKGGKGGKVGGAK21
M/Z: 621.3515, Charge: 2+, Mass Error: 0ppm, Ion Score: 55
WT_2_G_1 #925-1003 RT: 27.36-28.85 AV: 5 NL: 1.50E3T: Average spectrum MS2 621.35 (925-1003)
200 300 400 500 600 700 800 900 1000 1100 1200m/z
0
10
20
30
40
50
60
70
80
90
100
Rel
ativ
e A
bund
ance
999.01
715.04
612.08
526.98
910.11942.09
331.90
1095.05431.02 811.04
658.05284.881112.99509.06
1024.06356.96 793.03260.95 469.95 835.08 1201.44
y5
b5
y8
b13
b4
y7
y3
y10
b6
b9
b12
y9
y4
y12,2+
b10
y11
b11
[M-H2O+2H]2+
H2A.Z-K13AcK16AcK21Ac8GGKGGKGGKVGGAKNK23
M/Z: 774.3933, Charge: 2+, Mass Error: 0ppm, Ion Score: 102
WT_2_G_1 #1184 RT: 33.55 AV: 1 NL: 1.09E3T: ITMS + c NSI d Full ms2 [email protected] [200.00-1560.00]
200 400 600 800 1000 1200 1400m/z
0
10
20
30
40
50
60
70
80
90
100
Rel
ativ
e A
bund
ance
625.00
764.86 923.11
1012.99
755.89
715.99 1301.08535.00
1111.00707.13 822.95
567.08 954.79288.99 897.04
437.08 1211.021398.82
360.93 516.97
1417.00y2
b5
y8
b13
b4
y7
y3
y10
b6
b9
b12
y9
y11,2+
b10
y11
b11
[M-H2O+2H]2+
y6
y12
y13
y13,2+
y14,2+
b14
b15
H2A.Z-K10AcK13AcK16AcK21Ac8GGKGGKGGKVGGAKNK23
M/Z: 784.4318, Charge: 2+, Mass Error: 1ppm, Ion Score: 81
WT_2_G_1 #1440-1442 RT: 39.58-39.62 AV: 2 NL: 1.19E3T: Average spectrum MS2 784.43 (1440-1442)
400 600 800 1000 1200 1400m/z
0
10
20
30
40
50
60
70
80
90
100
Rel
ativ
e A
bund
ance
999.05
715.16 1236.04
1283.12835.05942.13569.03
1218.04853.06775.06331.94 431.00 1137.08550.96 1421.22642.08 1119.15
884.97284.98
1402.97415.92 1066.11511.69 1439.05b5
y8
b13
b3
y7
y3
y10
b6
b9
y9
y12,2+
b10
y11
b11
[M-H2O+2H]2+
y6
y12
y13
y13,2+
y14,2+b14
b15
y4 y5b4
H2A.Z-K16AcK21Ac11GGKGGKVGGAKNK23
M/Z: 621.351, Charge: 2+, Mass Error: -1ppm, Ion Score: 63
WT_2_G_1 #925-1003 RT: 27.36-28.85 AV: 5 NL: 1.50E3T: Average spectrum MS2 621.35 (925-1003)
200 300 400 500 600 700 800 900 1000 1100 1200m/z
0
10
20
30
40
50
60
70
80
90
100
Rel
ativ
e A
bund
ance
999.01
715.04
612.08
526.98
910.11942.09
331.90
1095.05431.02 811.04
658.05284.881112.99509.06
1024.06356.96 793.03260.95 469.95 835.08 1201.44
b5
y8
b12
b3
y7
y3
y10
b6
b9
y9
y11,2+
b10
y11
b11
[M-H2O+2H]2+
y4
y5y2
M/Z: 725.3801, Charge: 1+, Mass Error: 0ppm, Ion Score: 31
WT_3_G_3 #1102 RT: 32.03 AV: 1 NL: 1.44E3T: ITMS + c NSI d Full ms2 [email protected] [185.00-740.00]
200 250 300 350 400 450 500 550 600 650 700m/z
0
10
20
30
40
50
60
70
80
90
100
Rel
ativ
e A
bund
ance
558.98
337.03
437.02
288.98 576.93459.01 707.02
608.95388.79 531.21504.84278.98
360.86 653.01304.91 696.59270.98 729.03220.79
b4
b3
y6
y3
b7
y4
y5
y2
b7-H2O
H2A.Z-K16Ac14GGKVGGAK21
M/Z: 494.7563, Charge: 2+, Mass Error: 0ppm, Ion Score: 66
H2A.Z-K140Ac137SSAKTAEPR145
WT_3_G_3 #970 RT: 29.01 AV: 1 NL: 3.68E3T: ITMS + c NSI d Full ms2 [email protected] [125.00-1000.00]
200 300 400 500 600 700 800 900 1000m/z
0
10
20
30
40
50
60
70
80
90
100
Rel
ativ
e A
bund
ance
485.43
271.91
572.98
716.99476.45
400.92698.96407.45
743.07587.75415.79 814.06681.16551.88372.02253.97174.95 796.11 837.10616.13 926.29307.97
b4
b3
y6
y3
b7
y4
y5
y2
b7-H2O
y1
y7
y7,2+
[M-H2O+2H]2+
b8
H2B-1-A1Me1K3Me11APKKAPAAAAEK12
M/Z: 682.373, Charge: 2+, Mass Error: 0ppm, Ion Score: 52
E2+T1_2_H2B_G_2 #3108 RT: 50.31 AV: 1 NL: 3.88E2T: ITMS + c NSI d Full ms2 [email protected] [175.00-1375.00]
200 300 400 500 600 700 800 900 1000 1100 1200 1300m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100R
ela
tive
Ab
un
da
nce
1159.58
957.59 1029.60
885.52
479.22
664.23
813.51
673.33
567.92335.11
407.30
317.29
205.16 514.98
700.231176.67
793.311074.60243.12
719.89
y4
y5
y1
y7
b8
b7
b11
b9b10
y2y3
y8
y9
y2-NH3
b6,2+
b11,2+
[M-2H2O+2H]2+
[M-H2O+2H]2+
H2B-1-K3Me21APKKAPAAAAEK12
M/Z: 646.8825, Charge: 2+, Mass Error: 0ppm, Ion Score: 34
WT2_H2B_propionylated_2 #1698 RT: 34.99 AV: 1 NL: 4.64E2T: ITMS + c NSI d Full ms2 [email protected] [165.00-1305.00]
200 300 400 500 600 700 800 900 1000 1100 1200 1300m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
1193.59
438.33628.22
946.50473.85 636.41 804.58598.09347.31 1146.29728.56 875.41 1017.49258.19847.35779.75 y11
b8b7
b11b9
b10y3
y8
y9
y2-H2O
b8,2+
b9,2+
b2b3-NH3
b5
H2B-1-K3Me31APKKAPAAAAEK12
M/Z: 653.8902, Charge: 2+, Mass Error: 0ppm, Ion Score: 56
WT2_H2B_propionylated_2 #1706 RT: 35.11 AV: 1 NL: 4.26E2T: ITMS + c NSI d Full ms2 [email protected] [170.00-1320.00]
200 300 400 500 600 700 800 900 1000 1100 1200 1300m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
409.77
445.27
960.46
1031.59
889.62
1160.66
1207.56818.61
480.68
396.16 645.12579.54347.10
258.23 489.35657.69325.76 861.76222.20 539.55 728.40 1132.30985.81931.32
y5
y7
b8
b7
b11
b9
b10
y2
y3
y8b6,2+
b8,2+
y2-H2Ob4
b7,2+
b9,2+
[M-H2O+2H]2+
WT2_H2B_propionylated_1 #1727-1732 RT: 38.84-38.90 AV: 3 NL: 5.49E2T: Average spectrum MS2 731.93 (1727-1732)
200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
689.60
709.55
841.46912.43
641.04622.50
551.38
1096.52
1149.65
1423.86331.21 723.09466.61 932.33
790.46369.33 1280.691003.65 1377.841219.77237.35
H2B-1-A1Me11APKKAPAAAAEKK13
M/Z: 731.9352, Charge: 2+, Mass Error: 0ppm, Ion Score: 47
y4y5 b7
b3
b9
b10
y2
y3
y8y9
b5
b9,2+
y10
y11y12
y12,2+
y11,2+
b4
b11
[M-H2O+2H]2+
H2B-1-K3Me3K4Ac1APKKAPAAAAEKK13
M/Z: 738.9436, Charge: 2+, Mass Error: 1ppm, Ion Score: 53
WT2_H2B_propionylated_3 #1911 RT: 38.29 AV: 1 NL: 3.68E2T: ITMS + c NSI d Full ms2 [email protected] [190.00-1490.00]
200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
1146.66
1163.69
946.56
1017.65875.61
438.26
331.371331.75
721.54608.42 804.59
673.72
531.46 1348.89730.21402.72
296.231119.85
776.66268.35 1377.861221.10
y4
y5
b7
b9
b10
y2
y6
y8y9
b8
b7,2+
y10
b11
y3-H2O
y12-H2O
b6,2+
b8,2+
[M-2H2O+2H]2+
H2B-1-K115Ac104HAISEGTKAVTKFSSSTN121
M/Z: 1010.0059, Charge: 2+, Mass Error: 1ppm, Ion Score: 40
WT2_H2B_propionylated_3 #3250-8662 RT: 51.98-99.34 AV: 70 NL: 4.95E2T: Average spectrum MS2 1010.02 (3250-8662)
400 600 800 1000 1200 1400 1600 1800 2000m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
1106.46
991.95
1538.61
1391.54
1373.52
1625.661207.45936.37
1641.58913.38628.30 1425.54877.34 1825.671267.53481.25 769.87592.29378.18 1979.65
y17
y7
y16,2+
y10
y8
y12
b8
b7
b11
y14-NH3
b6
b10
y11
y13
y14
y15-NH3
y15
[M-2H2O+2H]2+
WT3_H2B_propionylated_2 #3593 RT: 57.39 AV: 1 NL: 2.78E3T: ITMS + c NSI d Full ms2 [email protected] [270.00-2000.00]
400 600 800 1000 1200 1400 1600 1800 2000m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Re
lativ
e A
bu
nd
an
ce
1011.57
954.181106.48789.17
1463.78 1679.73723.82 1373.60857.89666.41519.29 1593.65 1846.07378.06 1695.761207.33 1947.50
y8y10 y11
b3y13
y14y15
y16 y17y7,+2 y13,+2y17,+2
b3-H2O
b10
b11
[M-H2O+2H]2+
H2B-1-SSST(117-120)Phos104HAISEGTKAVTKFSSSTN121
M/Z: 1028.9841, Charge: 2+, Mass Error: 1ppm, Ion Score: 38
H2B-2-K3Me31APKKAPAATTEK12
M/Z: 719.3925, Charge: 2+, Mass Error: 1ppm, Ion Score: 44
WT2_H2B_propionylated_3 #2105-3280 RT: 40.91-52.25 AV: 4 NL: 2.69E2T: Average spectrum MS2 719.39 (2105-3280)
200 400 600 800 1000 1200 1400 1600 1800 2000m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
701.36
1233.70
1001.60657.39 1103.60
899.53611.13 827.50
539.25 781.46437.29335.24
1215.541262.52
y7
b8b7
b11
b10
y1
y2 y3y4
y5
b5
b9
[M-2H2O+2H]2+
H2B-2-A1Me11APKKAPAATTEKK13
M/Z: 761.9465, Charge: 2+, Mass Error: 1ppm, Ion Score: 54
WT2_H2B_propionylated_3 #1854-5129 RT: 37.75-67.71 AV: 9 NL: 1.89E2T: Average spectrum MS2 761.95 (1854-5129)
200 400 600 800 1000 1200 1400 1600 1800 2000m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100R
elat
ive
Abu
ndan
ce719.59
744.31
901.48
972.46
622.35
1156.53551.36331.31
1192.58384.16466.27 829.30 1340.61
282.18 1423.891045.221657.711291.56
y7
b8b7b3
b10
y2
y8
y5
b5
b9
y9
y10b4
b6
b11
b12
H2B-2-K3Me21APKKAPAATTEKK13
M/Z: 768.9542, Charge: 2+, Mass Error: 1ppm, Ion Score: 49
WT2_H2B_propionylated_3 #1771-4369 RT: 36.51-61.71 AV: 7 NL: 1.25E2T: Average spectrum MS2 768.95 (1771-4369)
200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
1206.66
1223.74751.43
760.06
875.60
331.28976.49
1390.73636.31 1077.64582.92804.45
561.45677.89
506.37 1178.62402.71 901.481033.76 1409.011161.72313.28 1372.74 1479.99
y7
b8
b7,2+
b10
y2
y8
y5
b5
b9
y9y12
b11
b2
y3
y4
b12
[M-H2O+2H]2+
[M-2H2O+2H]2+
H2B-2-K3Me31APKKAPAATTEKK13
M/Z: 784.4372, Charge: 2+, Mass Error: 1ppm, Ion Score: 58
WT2_H2B_propionylated_3 #1852-1858 RT: 37.74-37.81 AV: 3 NL: 4.38E2T: Average spectrum MS2 784.44 (1852-1858)
300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
1233.73
1251.75775.68
899.52
335.34
766.451001.58
1419.79657.38
1103.50827.62
911.48414.16 669.40567.53
983.46317.31 447.271437.711215.55525.51 1059.55597.47299.27
1392.80
b8
b8,2+
b10
y2
y8b3
b9
y9
b11
y4
b12
b5
b6
b7
b11-NH3
y2-NH3 y3-NH3
H2B-2-K3Me3K4Ac1APKKAPAATTEKK13
M/Z: 777.4291, Charge: 2+, Mass Error: 1ppm, Ion Score: 51
WT2_H2B_propionylated_3 #1762-2966 RT: 36.33-49.41 AV: 5 NL: 2.00E2T: Average spectrum MS2 777.43 (1762-2966)
200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
1219.61
1237.41
768.47
759.41
335.25
885.551089.69
987.60
1405.71
643.44407.26
317.31567.25
813.00 911.54545.32
443.22 1045.741191.74
1423.84286.45 1155.42 1342.64 1515.88
b8
b8,2+
b10
y2
y8
b9
y9
b11
y4
b12
b5
b11-NH3
y2-NH3
y3-NH3
y3
y5
y6,2+
y7,2+
y7
y10
[M-H2O+2H]2+
[M-2H2O+2H]2+
H3-K4Me13TKQTAR8
M/Z: 387.7271, Charge: 2+, Mass Error: 0ppm, Ion Score: 33
WT3_2 #3478 RT: 63.43 AV: 1 NL: 1.15E3T: ITMS + c NSI d Full ms2 [email protected] [95.00-790.00]
100 150 200 250 300 350 400 450 500 550 600 650 700 750m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
378.74
475.37
369.12
365.12
309.13
175.04 347.39227.06 295.02
265.18 428.29 600.34127.15 489.25 582.31556.30413.43 617.43 654.85202.96 683.58
b3
y1y3
b1
y4
b2
y5y2
[M-2H2O+2H]2+
y4-H2O
y5,2+
y5-NH3
[M-H2O+2H]2+
H3-K4Me23TKQTAR8
M/Z: 400.2186, Charge: 2+, Mass Error: 0ppm, Ion Score: 23
WT1_2 #2243-2937 RT: 42.82-56.18 AV: 3 NL: 5.32E2T: Average spectrum MS2 400.22 (2243-2937)
100 150 200 250 300 350 400 450 500 550 600 650 700 750 800m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
391.16
378.17
621.32
369.17179.16
321.26
483.33549.19
339.05265.08 403.08 603.39161.11475.32255.04
531.35205.21 419.41 641.44505.35140.10
691.60 745.34
b3
y1
y3
y4
b2
y5
y2
[M-2H2O+2H]2+
y5,2+
[M-H2O+2H]2+
b4
b5
b5-H2O
H3-K4Me33TKQTAR8
M/Z: 401.743, Charge: 2+, Mass Error: 1ppm, Ion Score: 19
WT1_2 #2188-2206 RT: 41.87-42.19 AV: 3 NL: 3.98E2T: Average spectrum MS2 401.74 (2188-2206)
100 150 200 250 300 350 400 450 500 550 600 650 700 750 800m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
628.35
379.96
392.70
175.18557.28
158.14600.49
287.07371.58 584.55 646.31456.35270.25
402.86551.07246.27 354.31 513.34139.19 428.39211.25 743.51710.37
b3
y1
y3
y4b2
y5
y2
[M-2H2O+2H]2+
[M-H2O+2H]2+
b4
b1
y5-NH3
H3-K9Ac9KSTGAKAPR17
M/Z: 535.3029, Charge: 2+, Mass Error: -1ppm, Ion Score: 61
WT1_2 #3186-5769 RT: 60.68-81.62 AV: 29 NL: 6.81E3T: Average spectrum MS2 535.28 (3186-5769)
200 300 400 500 600 700 800 900 1000m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100R
elat
ive
Abu
ndan
ce526.29
517.30
829.47
843.44
798.24
343.27272.20727.22241.15
227.07
742.47641.34277.20 598.40397.19 709.28215.19 471.96 894.62 981.56 1026.46b3y1
y3
b6y5
y2
b8
y6y7
b2
b1
b4b7
y8
[M-H2O+2H]2+
[M-2H2O+2H]2+
y4
b5
H3-K14Ac9KSTGAKAPR17
M/Z: 507.2902, Charge: 2+, Mass Error: -1ppm, Ion Score: 35
WT3_3 #3469-7674 RT: 63.41-99.81 AV: 12 NL: 1.80E3T: Average spectrum MS2 507.26 (3469-7674)
200 300 400 500 600 700 800 900 1000m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
489.25
321.09
498.33
471.20
742.38272.20 432.97343.21394.96
756.47724.32641.45 829.46226.20
527.39 584.35 787.55 839.41175.11 992.50885.43b3
y1
y3
y4
b6y5
y2
[M-2H2O+2H]2+
[M-H2O+2H]2+
b8
y6
y7
y8y7-H2O
H3-K9AcK14Ac9KSTGAKAPR17
M/Z: 528.2957, Charge: 2+, Mass Error: 0ppm, Ion Score: 60
WT1_2 #3378 RT: 62.69 AV: 1 NL: 4.50E3T: ITMS + c NSI d Full ms2 [email protected] [135.00-1070.00]
200 300 400 500 600 700 800 900 1000m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100R
elat
ive
Abu
ndan
ce519.32
829.45
510.31
784.41227.24 343.28272.18
713.06
641.48742.46
314.26 415.24 543.02 584.54454.18 695.30353.21175.17 881.36
b3y1
y3
b6
y5
y2
b8
y6y7
y8[M-H2O+2H]2+
[M-2H2O+2H]2+
b2
b4,2+
b5
b7
H3-K9AcK14Me19KSTGAKAPR17
M/Z: 549.2891, Charge: 2+, Mass Error: -3ppm, Ion Score: 31
[M-H2O+2H]2+WT2_1 #3613 RT: 63.51 AV: 1 NL: 1.98E3T: ITMS + c NSI d Full ms2 [email protected] [140.00-1110.00]
200 300 400 500 600 700 800 900 1000 1100m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
540.00
531.22
854.47821.10
869.50
749.18
277.15243.08 348.40521.33
435.00229.10 489.31 803.17576.97295.29 621.33 679.95404.46 919.38215.21 970.30 1076.75
[M-2H2O+2H]2+
y1
y3
b6
y5
y2
b8y6
y7b2
b1
b7
y8
y8,2+
H3-K18Ac18KQLASKAAR26
M/Z: 563.8324, Charge: 2+, Mass Error: 0ppm, Ion Score: 57
WT1_2 #6014 RT: 83.40 AV: 1 NL: 4.58E4T: ITMS + c NSI d Full ms2 [email protected] [145.00-1140.00]
200 300 400 500 600 700 800 900 1000 1100m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
881.20
900.53
952.19
246.23 317.25792.14
554.83 588.54539.17
450.86227.13
501.45 645.52175.03 772.56
443.87810.11
868.43355.35260.29 740.61 971.98
y1
y3
b6
y5
y2
b8
y6
y7
b2
b1
b7
y8
y8,2+
y4
y7-H2O
b4
b5
b6-H2O
H3-K23Ac18KQLASKAAR26
M/Z: 563.8318, Charge: 2+, Mass Error: -1ppm, Ion Score: 54
WT1_1 #5983 RT: 83.14 AV: 1 NL: 4.47E4T: ITMS + c NSI d Full ms2 [email protected] [145.00-1140.00]
200 300 400 500 600 700 800 900 1000 1100m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100R
elat
ive
Abu
ndan
ce881.19
900.58
952.35
317.22246.25
792.08
588.42
574.35443.72227.07
659.42758.52450.77
175.19
809.97
369.05282.19 740.51408.03 1067.80
y1
y3
b6
y5
y2
b8
y6y7
b2
b1
b7
y8
y8,2+
y4
y7-H2O
b3
b4
b5
b6-H2O
H3-K18AcK23Ac18KQLASKAAR26
M/Z: 556.8272, Charge: 2+, Mass Error: 4ppm, Ion Score: 51
WT1_2 #3382-10333 RT: 62.74-118.45 AV: 25 NL: 2.48E3T: Average spectrum MS2 557.26 (3382-10333)
200 300 400 500 600 700 800 900 1000 1100m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
886.54
867.21
938.28539.19
547.93
246.15 574.42317.26227.16
778.14443.81
744.56645.43417.31468.19
175.15 637.48789.48
395.34 796.301081.53367.24
272.27 727.53 840.39 1063.52957.52 1092.44
y1
y3
b6
y5y2
b8
y6
y7
b2
b1
b7
y8
y8,2+
y4
b3
b4
b5
b6-H2O
[M-H2O+2H]2+
H3-K23Me118KQLASKAAR26
M/Z: 585.3263, Charge: 2+, Mass Error: 1ppm, Ion Score: 55
WT1_2 #5230-10383 RT: 77.55-118.98 AV: 48 NL: 7.26E3T: Average spectrum MS2 585.27 (5230-10383)
200 300 400 500 600 700 800 900 1000 1100m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
927.50
576.24919.28243.07
991.25
611.39323.23 829.20559.12464.18251.19
683.45
909.51509.36 797.48661.17
455.24179.07 373.12 847.12
437.28 779.43305.21215.05 1009.59 1130.65
y1
y3
b6
y5
y2
b8
y6
y7
b2
b1 b7
y8
y8,2+
y4
y7-H2O
b4 b6-NH3
y8-NH3
[M-H2O+2H]2+
H3-K23Me218KQLASKAAR26
M/Z: 556.842, Charge: 2+, Mass Error: -2ppm, Ion Score: 35
WT3_3 #5307 RT: 78.88 AV: 1 NL: 2.46E3T: ITMS + c NSI d Full ms2 [email protected] [140.00-1125.00]
200 300 400 500 600 700 800 900 1000 1100m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
872.52
547.75
241.05 744.58
631.50538.39 938.40436.79
428.13
855.53
482.11796.43
317.26 372.91175.16727.80560.47
229.11 956.56896.53 1082.59281.16 684.41962.84
y1 y3b6
y5
y2
b8
y6
y7
b2
b1
b7
y8
y7,2+
y4
b3
y7-NH3
[M-H2O+2H]2+
y2-NH3
y8-NH3
y8,2+
H3-K23Me318KQLASKAAR26
M/Z: 563.8512, Charge: 2+, Mass Error: 1ppm, Ion Score: 22
WT1_2 #5008 RT: 75.87 AV: 1 NL: 2.02E4T: ITMS + c NSI d Full ms2 [email protected] [145.00-1140.00]
200 300 400 500 600 700 800 900 1000 1100m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
555.17
885.53
952.44
242.91
641.39434.15
867.48545.84 628.44 741.48
378.29323.33 970.58586.46175.09 482.30251.17810.69699.51 924.63
229.03 276.451024.77
y1
y3
b6
b8
y6
y7
b2
b1
y6,2+b3
[M-H2O+2H]2+
y8-NH3
y8,2+y6-NH3
y7,2+
b5
H3-K23Form18KQLASKAAR26
M/Z: 556.8272, Charge: 2+, Mass Error: 4ppm, Ion Score: 25
WT1_2 #5711 RT: 81.19 AV: 1 NL: 1.89E4T: ITMS + c NSI d Full ms2 [email protected] [140.00-1125.00]
200 300 400 500 600 700 800 900 1000 1100m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
886.49
867.16
938.29
539.13246.12
443.88317.19 574.49
227.11 645.36
637.64778.34
487.42175.14 796.13626.28367.21
395.14 744.55 850.64265.08 1092.361004.70
y1
y3
b6-H2O
b8
y6y7
b1
[M-H2O+2H]2+
y8,2+
b7
y2
y8
H3-K27AcK36Me227KSAPATGGIKKPHR40
M/Z: 815.4768, Charge: 2+, Mass Error: 2ppm, Ion Score: 75
WT1_2 #4408 RT: 70.80 AV: 1 NL: 4.40E2T: ITMS + c NSI d Full ms2 [email protected] [210.00-1645.00]
300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 1600m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
623.71
797.43
702.651245.72
1221.70
816.57
409.31
771.551403.53
1077.53511.44976.46593.33
1317.011148.83314.07296.20 488.91 837.43368.25
1474.80919.57
y3
b2-H2O
b13
y6y7
b2
y13,2+
b10
y2
y8y4
y9
y10
y11
y12
y13y12,2+
y11,2+
y9,2+y8,2+b3-NH3
b11
b10-NH3
b13-H2O
H3-K27Ac27KSAPATGGIKKPHR40
M/Z: 829.4732, Charge: 2+, Mass Error: 0ppm, Ion Score: 71
WT1_2 #5076 RT: 76.42 AV: 1 NL: 1.83E3T: ITMS + c NSI d Full ms2 [email protected] [215.00-1670.00]
300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 1600m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
637.421273.63
409.35
716.37
1004.64
820.371431.64
1105.59593.491249.52
865.53 947.49
1483.541023.32540.30 1403.69367.05 1176.64296.19 513.29 1344.25
y3
b2-H2O
b13
y6
y7
b2
y13,2+
b10
y8
y4 y9
y10
y11
y12
y13
y12,2+
y11,2+
y9,2+
b11
[M-H2O+2H]2+
y5
b3-H2O
b9
H3-K27Me127KSAPATGGIKKPHR40
M/Z: 843.489, Charge: 2+, Mass Error: 1ppm, Ion Score: 90
WT2_3 #6077 RT: 81.86 AV: 1 NL: 1.82E4T: ITMS + c NSI d Full ms2 [email protected] [220.00-1700.00]
400 600 800 1000 1200 1400 1600m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
1273.62
637.36
409.28
1431.65
716.40
834.561004.49593.34
1105.60947.54395.15 1511.63865.34
255.221344.70
553.35497.39324.20 1176.521529.17
y3
b2-H2O
b13
y6
y7
b3
y13,2+
b10
y8y4
y9
y10
y11
y12
y13
y12,2+
y11,2+
y9,2+
b11
[M-H2O+2H]2+
y5
b3-H2O
b9
b1
b4
H3-K27Me227KSAPATGGIKKPHR40
M/Z: 822.4842, Charge: 2+, Mass Error: 1ppm, Ion Score: 88
WT2_3 #4437-5173 RT: 68.10-74.78 AV: 11 NL: 9.54E2T: Average spectrum MS2 822.48 (4437-5173)
300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 1600m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100R
elat
ive
Abu
ndan
ce813.63
1273.65
1235.57804.57
409.30
1431.681004.57792.72371.26 1469.65
593.401105.59 1344.62
623.46343.27 867.31539.32 947.66702.49 1176.56
468.39 1487.551584.71
[M-H2O+2H]2+
y3
b4-NH3
b13
y6
y7
b3
b10
y8
y4y9
y10
y11
y12
y13
b11
b5
[M-2H2O+2H]2+
H3-K27Me327KSAPATGGIKKPHR40
M/Z: 829.4902, Charge: 2+, Mass Error: -1ppm, Ion Score: 38
WT2_3 #3661-5510 RT: 60.16-77.50 AV: 16 NL: 7.78E2T: Average spectrum MS2 829.45 (3661-5510)
300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 1600m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
1273.64
637.44
409.29811.43
820.50716.39
799.49 1004.55593.44
1431.691105.59830.52
1483.671249.54
522.25947.55367.13
1023.41494.12
1231.41254.12 1344.61
1501.731633.39
[M-H2O+2H]2+
y3
b13
y6
y7
b3
b10
y8y4
y9
y10
y11
y12
y13
b11
b6
[M-2H2O+2H]2+
y5b7 b8 b9
H3-K27Me1K36Me127KSAPATGGIKKPHR40
M/Z: 850.4978, Charge: 2+, Mass Error: 2ppm, Ion Score: 88
WT2_1 #5851 RT: 82.06 AV: 1 NL: 2.38E3T: ITMS + c NSI d Full ms2 [email protected] [220.00-1715.00]
400 600 800 1000 1200 1400 1600m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
1287.64
644.49
723.43409.311445.68
791.47593.421018.59
841.50
850.981119.61395.14
961.691525.53
879.52255.15 1358.79342.30
510.011190.58
1544.64 1676.55
[M-H2O+2H]2+
y3
b13
y6
y7
b10
y8
y4
y9
y10
y11
y12
y13
b1
[M-2H2O+2H]2+
y5
y2
y8,2+
y11,2+
y12,2+
y13,2+
b2
b11-H2O
H3-K27Me1R40Me127KSAPATGGIKKPHR40
M/Z: 850.4975, Charge: 2+, Mass Error: 1ppm, Ion Score: 99
WT2_2 #5785 RT: 80.26 AV: 1 NL: 9.30E2T: ITMS + c NSI d Full ms2 [email protected] [220.00-1715.00]
400 600 800 1000 1200 1400 1600m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
644.57
1287.66
1445.63
851.39723.24423.28
841.32832.30 1018.55
607.33791.64
961.67
255.14 1119.65395.09 560.37
865.51 1511.741259.38 1358.76
324.22 1525.461177.10542.74
1677.79
[M-H2O+2H]2+
y3
b13
y7
b9
y8
y4
y9
y10
y11
y12
y13
b1
[M-2H2O+2H]2+
y5
y2
y9,2+
y11,2+
y12,2+
y13,2+
b3b2-H2O
b3-H2O
H3-R40Me127KSAPATGGIKKPHR40
M/Z: 843.4823, Charge: 2+, Mass Error: -7ppm, Ion Score: 65
WT2_1 #5127 RT: 76.35 AV: 1 NL: 8.16E2T: ITMS + c NSI d Full ms2 [email protected] [220.00-1700.00]
400 600 800 1000 1200 1400 1600m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
1287.68
825.59644.32
843.94723.52
1018.67
799.56
423.41
607.44536.27 1445.701119.69
381.351497.62465.56 961.59
310.38 1245.22904.561427.791214.64 1511.68
y3
b13y7
b9
y8
y4
y9
y10
y11
y12
y13
b1
y5
y2
y11,2+
y12,2+
y13,2+
b3
b3-H2O
y6
b11
b11-H2O
H3-R49Me141FRPGTVALR49
M/Z: 551.3031, Charge: 2+, Mass Error: 1ppm, Ion Score: 41
WT2_1 #7002 RT: 91.03 AV: 1 NL: 4.73E3T: ITMS + c NSI d Full ms2 [email protected] [140.00-1115.00]
200 300 400 500 600 700 800 900 1000 1100m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
737.49
365.26
542.47440.30
533.17 795.41
723.31879.46639.40
175.20 379.30337.35 479.41289.25 551.18 695.41 925.58811.35 974.57 1082.90
[M-H2O+2H]2+
y7
b8
y6
b1
[M-2H2O+2H]2+
y5y2
b2
y1
y1-NH3 y2-NH3 y3-NH3
y8-NH3
y8-NH3,2+
b5
b6
b7
H3-K56Me153KYQKSTDLLIR63
M/Z: 754.4066, Charge: 2+, Mass Error: -1ppm, Ion Score: 47
WT2_1 #6565 RT: 87.66 AV: 1 NL: 4.87E2T: ITMS + c NSI d Full ms2 [email protected] [195.00-1520.00]
200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100R
elat
ive
Abu
ndan
ce736.34
745.58
841.41
1027.57
521.40
1157.58
570.17407.26 481.28667.32 1329.31
1101.11351.04
579.45987.26
755.38637.33293.15
1215.45
961.32826.941283.73228.41
1375.07
[M-H2O+2H]2+
y8
b8
y6
[M-2H2O+2H]2+
y5y2
b2
y3
y9-NH3
b9
b10
b7
y4
y9
y10
y9,2+
b3
H3-K79Me173DIAHEFKAELR83
M/Z: 727.8854, Charge: 2+, Mass Error: 0ppm, Ion Score: 41
WT2_3 #9463 RT: 107.87 AV: 1 NL: 6.42E3T: ITMS + c NSI d Full ms2 [email protected] [190.00-1470.00]
200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
642.43
550.39
962.57
833.42
1099.57
585.79
493.10 718.88 1167.27
953.31 1280.29431.31 769.321024.43288.24
944.50338.161298.021207.86 1434.16
[M-H2O+2H]2+
y8
b10
y6
y5y2
b2
b9
b6
y4y9
y10
y9,2+
b4
y7
y8,2+
y10,2+
b3-H2O
H3-R83Me173DIAHEFKAELR83
M/Z: 727.8854, Charge: 2+, Mass Error: 0ppm, Ion Score: 49
WT1_1 #9129 RT: 107.97 AV: 1 NL: 6.23E3T: ITMS + c NSI d Full ms2 [email protected] [190.00-1470.00]
200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
642.44
550.36
962.54
833.46
1099.50
718.90585.91
502.50 686.39 1167.27493.22 1280.53
819.56288.23 953.39 1024.39431.28 769.21
249.25 355.55 944.51 1262.14 1298.32
[M-H2O+2H]2+
y8
b8
y6
y5
y2b2
b7
b6
y4
y9
y9,2+
b4
y7
y8,2+
y10,2+
b2-H2O
y3
b3
b5
H3-R128Me1116RVTIMTKDMQLAR128
M/Z: 844.964, Charge: 2+, Mass Error: 1ppm, Ion Score: 46
WT1_3 #7738 RT: 98.04 AV: 1 NL: 8.94E2T: ITMS + c NSI d Full ms2 [email protected] [220.00-1700.00]
400 600 800 1000 1200 1400 1600m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
1057.55
632.38
845.54
822.99
409.281274.65
1163.42716.69
800.56 1476.67942.271316.57 1429.60619.62466.22 931.65
1257.14356.15 1401.81 1512.89255.11
y8
b8
y5
b7
b6y4
y9
b4
y7
y12
b4-NH3,2+
y3
b5y3-NH3
b9
b10b11
H3.3-K9AcK14Ac9KSTGVKAPR17
M/Z: 549.2891, Charge: 2+, Mass Error: -3ppm, Ion Score: 44
WT2_2 #3810 RT: 63.93 AV: 1 NL: 1.42E3T: ITMS + c NSI d Full ms2 [email protected] [140.00-1110.00]
200 300 400 500 600 700 800 900 1000 1100m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100R
elat
ive
Abu
ndan
ce540.36
531.16
869.49
854.40
821.30
348.48229.18 519.35277.28 749.23577.17
489.43435.02 781.44665.47317.13 401.14 595.47 898.99711.96189.11 1023.30
b3
y3b6
y5
y2
b8y6
y7
y8
[M-H2O+2H]2+
[M-2H2O+2H]2+
b2
b1
b5
b7
y4
y8,2+
b3-NH3
H3.3-K9Ac9KSTGVKAPR17
M/Z: 556.2984, Charge: 2+, Mass Error: 0ppm, Ion Score: 32
WT2_3 #4341 RT: 67.37 AV: 1 NL: 4.57E3T: ITMS + c NSI d Full ms2 [email protected] [140.00-1125.00]
200 300 400 500 600 700 800 900 1000 1100m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
538.21
547.38
883.54
835.20349.24 763.13229.10
277.27
669.43577.07416.26 795.44515.41693.49317.33 620.30 1081.52179.19 896.73401.26 952.58b3
y3 b6
y5
y2
b8
y6y7
y8
[M-H2O+2H]2+
[M-2H2O+2H]2+
b2
b1
b5
b7
y4
y8,2+y1
H3.3-K14Form9KSTGVKAPR17
M/Z: 542.3081, Charge: 2+, Mass Error: -6ppm, Ion Score: 64
WT2_2 #1990-5681 RT: 35.62-79.49 AV: 38 NL: 2.16E3T: Average spectrum MS2 541.90 (1990-5681)
200 300 400 500 600 700 800 900 1000m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
533.24
843.49524.32
241.12 427.92
812.25343.26272.22
741.30
807.40655.47468.17598.46 857.56328.24 411.17 723.23227.12 910.48 1039.57969.62
y3
b6
y5
y2
b8
y6
y7
y8
[M-H2O+2H]2+
[M-2H2O+2H]2+
b2
b1
b7
b3-H2O
b4-H2O
H3.3-K18AcK23Ac18KQLATKAAR26
M/Z: 563.8318, Charge: 2+, Mass Error: -1ppm, Ion Score: 59
WT2_1 #5831 RT: 81.91 AV: 1 NL: 1.06E4T: ITMS + c NSI d Full ms2 [email protected] [145.00-1140.00]
200 300 400 500 600 700 800 900 1000 1100m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100R
elat
ive
Abu
ndan
ce886.51
900.51
952.30
792.13246.13
539.12
317.20 574.49
645.37
443.75227.16 659.37487.35
772.47175.13
810.09
355.16 868.50434.89
377.38 741.55269.22 1013.54 1104.72
y1
y3
b6
y5
y2
b3
y6y7
b2
b1
b7
y8
y8,2+y4
b4
b5
b6-H2O
[M-H2O+2H]2+
b8
H3.3-K23Form18KQLATKAAR26
M/Z: 563.8318, Charge: 2+, Mass Error: -1ppm, Ion Score: 45
WT2_1 #5831 RT: 81.91 AV: 1 NL: 1.06E4T: ITMS + c NSI d Full ms2 [email protected] [145.00-1140.00]
200 300 400 500 600 700 800 900 1000 1100m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100R
elat
ive
Abu
ndan
ce886.51
900.51
952.30
792.13246.13
539.12
317.20 574.49
645.37
443.75227.16 659.37487.35
772.47175.13
810.09
355.16 868.50434.89
377.38 741.55269.22 1013.54 1104.72
y1
y3
b6
y5
y2
b3
y6
y7
b2
b1
b7
y8
y8,2+
y4
b4
b5
b6-H2O
b8
b8-NH3,2+
H3.3-K56Ac53KYQKTTDLLIR63
M/Z: 738.9251, Charge: 2+, Mass Error: 1ppm, Ion Score: 56
WT2_2 #6830 RT: 88.10 AV: 1 NL: 9.11E2T: ITMS + c NSI d Full ms2 [email protected] [190.00-1490.00]
200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
817.47
706.08 1129.58
730.25
1001.38
348.21
660.38 1302.55
1189.43514.43629.31
320.20 476.381112.58401.20
521.32963.35293.35
556.41781.53 837.14 1266.48
275.40 458.37 930.53247.42 1347.58
y8
b8
y6
y5
y2
b2
y3y9-NH3
b9
b10
b7
y4
y9
y10y9,2+
b3
b4
b6
H3.3-K79Me173DIAMEMKSDIR83
M/Z: 725.3299, Charge: 2+, Mass Error: 0ppm, Ion Score: 26
WT2_2 #7193-9451 RT: 90.77-108.84 AV: 6 NL: 7.35E2T: Average spectrum MS2 725.38 (7193-9451)
200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
651.40
707.33
609.42
716.37 953.12496.31 1270.06329.27 401.04 725.88 1041.10231.20 815.34 944.44 1150.31 1329.23 1411.51
[M-H2O+2H]2+
y8
b8y5
y2b3
b7
y4b4 y7y3
[M-2H2O+2H]2+
H4-K4Ac1AGGKGGK7
M/Z: 728.3949, Charge: 2+, Mass Error: 2ppm, Ion Score: 35
WT2_H4_propionylated_1 #2727 RT: 46.97 AV: 1 NL: 5.70E3T: ITMS + c NSI d Full ms2 [email protected] [190.00-740.00]
200 250 300 350 400 450 500 550 600 650 700m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
601.26
710.23
469.17 526.24
412.15317.26 487.32 544.31285.10 572.17 692.22399.26342.15 653.93613.86260.25 507.92 718.78445.98228.05
b6
y5
y2
b3
b5
y4b4
y6
y3
[M-2H2O+2H]2+
H4-K4AcK7Ac1AGGKGGKGMGK11
M/Z: 1143.5856, Charge: 1+, Mass Error: 2ppm, Ion Score: 64
WT2_H4_propionylated_1 #3940-3960 RT: 56.46-56.63 AV: 3 NL: 4.45E2T: Average spectrum MS2 1143.59 (3940-3960)
300 400 500 600 700 800 900 1000 1100m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
1125.36
1016.53
884.11
753.31
696.151099.39
1081.13941.21732.48 998.58855.91 1028.88525.43 675.25469.38 618.36 796.93 923.17417.24385.29
b6
y8
y5
b7
b5y9
b4
y6
y10
[M-2H2O+2H]2+
y7
b8
b9
b10
H4-K7Ac1AGGKGGKGMGK11
M/Z: 579.3083, Charge: 2+, Mass Error: 2ppm, Ion Score: 71
RIN1_2_H4_propionylated_3 #5141 RT: 61.29 AV: 1 NL: 2.40E3T: ITMS + c NSI d Full ms2 [email protected] [145.00-1170.00]
200 300 400 500 600 700 800 900 1000 1100m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Re
lativ
e A
bu
nd
an
ce
561.32
898.31
570.17
515.89
710.23506.78
732.39448.28
803.12260.30
767.05658.90
955.30
972.38391.30617.83 869.89
203.19 1030.33374.61
1117.98
y5
b7
b4
y6
y10
[M-2H2O+2H]2+
y7
b8
b9
b10
[M-H2O+2H]2+
y1
y2
y3
y4
y10,2+
H4-K4AcK7AcK11AcK15Ac1AGGKGGKGMGKVGAKR16
M/Z: 841.9559, Charge: 2+, Mass Error: 2ppm, Ion Score: 51
EZL2_1_H4_propionylated_1 #4110 RT: 58.92 AV: 1 NL: 1.85E3T: ITMS + c NSI d Full ms2 [email protected] [220.00-1695.00]
400 600 800 1000 1200 1400 1600m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
823.40
832.63778.49
657.28473.36 1210.37572.46 1111.17
1508.271338.24884.16317.15 1083.44393.24 1195.27941.05 1567.721377.35
y5
b7b4 y8 y10
[M-2H2O+2H]2+
y7
b9b10
[M-H2O+2H]2+
y2 y3
y4
y15,2+
y9 y11 y12b5
b11b12
b13
b14 b15
H4-K4AcK7AcK11Ac1AGGKGGKGMGKVGAKR16
M/Z: 848.9645, Charge: 2+, Mass Error: 2ppm, Ion Score: 67
EZL2_1_H4_propionylated_3 #4495 RT: 61.98 AV: 1 NL: 3.11E3T: ITMS + c NSI d Full ms2 [email protected] [220.00-1710.00]
400 600 800 1000 1200 1400 1600m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
785.41
839.94
1224.31
487.46
473.24
1522.591125.211001.48572.57586.55 848.57
756.95
696.26 1083.59 1285.60345.33 987.58 1352.33412.20 643.34 955.09 1395.75299.23
1565.96
[M-H2O+2H]2+
y5
b7
b4
y8y7
b9
b10
y2
y4
y15,2+
y9
y11
y12
b5
b11
b12
b13b14
b15
y6
b8
H4-K4AcK11Ac1AGGKGGKGMGKVGAKR16
M/Z: 855.9704, Charge: 2+, Mass Error: 1ppm, Ion Score: 68
EZL2_1_H4_propionylated_1 #4622-10907 RT: 62.91-119.51 AV: 7 NL: 6.49E2T: Average spectrum MS2 855.97 (4622-10907)
400 600 800 1000 1200 1400 1600m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
837.30
792.30
846.63
1224.49
487.35
1125.31633.20 1536.40521.21 1238.19651.19
473.361001.61 1352.43783.30435.30
1139.31710.22 884.11359.30 1285.86 1409.58958.23 1107.49277.11 1596.21
1671.911487.55
[M-H2O+2H]2+
y5
b6
y8
y7
b9
b10
y2
y4
y15,2+
y9
y11y12
b5
b11
b12
b13
b14
b15
b7
[M-2H2O+2H]2+
y3y14,2+
y13,2+
y12,2+
y9,2+
y8,2+
H4-K7AcK11Ac1AGGKGGKGMGKVGAKR16
M/Z: 855.9723, Charge: 2+, Mass Error: 3ppm, Ion Score: 62
WT3_H4_propionylated_3 #4764 RT: 62.08 AV: 1 NL: 1.99E3T: ITMS + c NSI d Full ms2 [email protected] [225.00-1725.00]
400 600 800 1000 1200 1400 1600m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
792.45
1224.42846.60487.37
1125.36586.42
1238.37
473.37569.38 1536.58
783.54651.35 898.241015.55767.29
1139.32 1352.40426.74 1097.52 1299.66
912.20345.30 1516.25307.52
1583.32
y5
b6y8
y7
b9
b10y2
y4
y15,2+
y9
y12
b4
b11
b12
b13
b14
b15
b7
[M-H2O+2H]2+
y3
y13,2+
y8,2+
y6
b8
[M-2H2O+2H]2+
H4-K11Ac1AGGKGGKGMGKVGAKR16
M/Z: 862.9801, Charge: 2+, Mass Error: 3ppm, Ion Score: 59
WT3_H4_propionylated_2 #5039 RT: 63.52 AV: 1 NL: 1.09E3T: ITMS + c NSI d Full ms2 [email protected] [225.00-1740.00]
400 600 800 1000 1200 1400 1600m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
799.21
845.49
1238.33
486.37
853.43
1550.66790.54 1139.26
469.271252.23585.53 781.35
1299.63912.25
1014.65430.62 572.42 625.39 1532.49359.371686.33
y5
y8
b9
y4
y15,2+
y9
y12
b4
b11
b12
b14
b15
b7
[M-H2O+2H]2+
y3y8,2+
b8
[M-2H2O+2H]2+
H4-K7Ub1AGGKGGKGMGKVGAKR16
M/Z: 870.9785, Charge: 2+, Mass Error: -2ppm, Ion Score: 41
RIN1_2_H4_propionylated_3 #5025 RT: 60.41 AV: 1 NL: 5.16E3T: ITMS + c NSI d Full ms2 [email protected] [225.00-1755.00]
400 600 800 1000 1200 1400 1600m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Re
lativ
e A
bu
nd
an
ce
807.41
1254.46487.37
853.30
586.33
1031.681566.581155.44 1315.72
1127.47711.38 1382.28914.39359.27 665.15329.26 1439.58 1611.57b6
y5
b7,2+b5
y12
b4
y15,2+
y11 b13
b12
b11
y2
y3
y4
b14
b15
b10
H4-K7AcK11AcK15Ac5GGKGMGKVGAKR16
M/Z: 1327.7186, Charge: 1+, Mass Error: 3ppm, Ion Score: 66
WT2_H4_propionylated_3 #4234 RT: 58.19 AV: 1 NL: 1.56E3T: ITMS + c NSI d Full ms2 [email protected] [355.00-1340.00]
400 500 600 700 800 900 1000 1100 1200 1300m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
1309.67
398.13756.05 837.26 1285.67
529.09782.53 987.46586.20 1197.31738.06 970.41
625.41 855.15 1157.68473.38724.51 1094.451022.56
b6
y5
b5
y11
b4
y6
b8
y3
y4
b7
y7
y8
y9
y10
[M-2H2O+2H]2+
y5-NH3
y7-NH3
y9-NH3
y11-NH3
H4-K11AcK15Ac8GMGKVGAKR16
M/Z: 1043.5687, Charge: 1+, Mass Error: 2ppm, Ion Score: 30
WT3_H4_propionylated_2 #4123 RT: 56.60 AV: 1 NL: 2.46E3T: ITMS + c NSI d Full ms2 [email protected] [275.00-1055.00]
300 400 500 600 700 800 900 1000m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
1025.55
999.50
472.14 628.12
571.23857.47
555.45 979.57456.39839.45782.51319.03 883.33610.33
359.26 681.41 930.70398.26 742.63512.40
653.25
b6
y5
b5
b4
y6b8
y3
b7
y7 y8
b3b4-NH3
b5-NH3
[M-2H2O+2H]2+
y4-NH3
y5-NH3
y6-NH3
y7-NH3
y8-NH3
H4-K15Ac8GMGKVGAKR16
M/Z: 529.2958, Charge: 2+, Mass Error: 2ppm, Ion Score: 41
WT3_H4_propionylated_1 #4932 RT: 60.53 AV: 1 NL: 1.83E4T: ITMS + c NSI d Full ms2 [email protected] [135.00-1070.00]
200 300 400 500 600 700 800 900 1000m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
473.19
487.37
883.27
585.10
520.32
586.40
472.03
407.24175.24713.06345.28
416.31 628.08317.24
813.68756.55
302.14203.17812.98245.19 815.47657.06 900.53 936.59
y5
b5
b4
y6
b8
y3
b7
y7
b3
[M-H2O+2H]2+
y4
y7,2+y1
y2
y8,2+
b2
H4-R35Me124ASIEGITKPAIR35
M/Z: 1397.761, Charge: 1+, Mass Error: 3ppm, Ion Score: 63
WT2_H4_propionylated_3 #7460 RT: 82.68 AV: 1 NL: 1.32E3T: ITMS + c NSI d Full ms2 [email protected] [370.00-1410.00]
400 500 600 700 800 900 1000 1100 1200 1300 1400m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Re
lativ
e A
bu
nd
an
ce
937.44
1379.74
1353.70
459.24
518.88919.67645.23 1181.47 1251.68717.46 1067.85765.81 857.44441.20 627.53 1002.88
y5
b5
y6y11
b7-H2O
y7
[M-H2O+2H]2+
y4y9
y10
y8
y4-NH3
y5-NH3
y6-NH3
y8-NH3 y11-NH3
H4-R55Me146ISSFIYDDSR55
M/Z: 1285.5776, Charge: 1+, Mass Error: 4ppm, Ion Score: 52
WT1_H4_propionylated_1 #7393 RT: 88.48 AV: 1 NL: 1.88E3T: ITMS + c NSI d Full ms2 [email protected] [340.00-1300.00]
400 500 600 700 800 900 1000 1100 1200 1300m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Re
lativ
e A
bu
nd
an
ce
1267.52
1255.47659.24
397.17609.19477.09
1241.56
1097.44939.38 1009.04773.42494.85 1115.60921.39 973.34677.25459.21 791.43 1184.98603.39y4
b5
y5
b4-H2O
y6
y3
y8y9
y7y9-H2O
[M-H2O+2H]2+
y8-H2O
y7-H2O
y6-H2O
y5-H2O
b4
b5
H4-R67Me156QVLKSFLENVVR67
M/Z: 1575.852, Charge: 1+, Mass Error: 3ppm, Ion Score: 54
WT1_H4_propionylated_1 #10257-10264 RT: 112.58-112.67 AV: 3 NL: 4.07E2T: Average spectrum MS2 1575.86 (10257-10264)
500 600 700 800 900 1000 1100 1200 1300 1400 1500m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Re
lativ
e A
bu
nd
an
ce
1557.80
1283.43
805.24657.12509.35
937.24 989.651297.43 1531.36679.35 1091.28491.41
1347.581157.75693.26 822.39
y4
b7
y5
b5-H2O
y6
y8
y9
y7
[M-H2O+2H]2+
y8-H2O
b4
b10
y10 y11
y4-NH3y9-NH3
b6-H2O
H4-R77Me168DAVTYTEHAR77
M/Z: 616.7969, Charge: 2+, Mass Error: -3ppm, Ion Score: 43
WT3_H4_propionylated_2 #4667 RT: 60.74 AV: 1 NL: 3.83E4T: ITMS + c NSI d Full ms2 [email protected] [155.00-1245.00]
200 300 400 500 600 700 800 900 1000 1100 1200m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Re
lativ
e A
bu
nd
an
ce
891.58
531.44
790.43
607.74
446.42
889.44
892.64495.62
443.15588.12 787.39 791.45
397.39342.11243.08888.49
314.31224.95 987.32689.13 1044.40887.48972.90
1175.30
[M-H2O+2H]2+
y4
b6
y5
b2-H2O
y6
y8 y9
y7
b4
b8
y3
y2
b6-H2Oy5,2+
y7,2+
y8,2+
y9,2+
b2 b3
b5-H2O
b9
b9,2+
H4-K91Me178RKTVTAMDVVYALKR92
M/Z: 967.0534, Charge: 2+, Mass Error: 3ppm, Ion Score: 76
WT3_H4_propionylated_2 #9725 RT: 100.37 AV: 1 NL: 1.24E3T: ITMS + c NSI d Full ms2 [email protected] [255.00-1945.00]
400 600 800 1000 1200 1400 1600 1800m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Re
lativ
e A
bu
nd
an
ce
918.691015.52
1336.651213.54
1033.59 1560.821447.80
1577.96 1721.42819.53720.51469.27
380.39558.34 1745.121716.89
1760.241631.02
[M-H2O+2H]2+
y4b6
y5y6
y8
y9
y7b8
b9
y2-NH3
y3-NH3
y10
y11
y12
y13
y14
b2-NH3
b10
b11
b12b13
b14
H4-R92Me179KTVTAMDVVYALKR92
M/Z: 889.0012, Charge: 2+, Mass Error: 1ppm, Ion Score: 43
WT3_H4_propionylated_1 #10705 RT: 105.45 AV: 1 NL: 5.47E2T: ITMS + c NSI d Full ms2 [email protected] [230.00-1790.00]
400 600 800 1000 1200 1400 1600m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100R
elat
ive
Abu
ndan
ce870.33
1291.18
1588.331057.15
557.59
720.55495.351220.11
1404.38486.34
958.19845.39
373.40 1336.78819.45
595.441536.90744.03
639.26 1164.81414.25 1602.691103.55
328.99 1504.94
[M-2H2O+2H]2+
y3
b6
y5
y6
y8y9
b8
b9
y11
y13
b10
b11
b12
b13
y2
y4
b5-H2O
b10,2+
b10-H2O
b11-H2O
b13-H2O
Figure S4. Down-regulation of H2B C-terminal Phosphorylation
HAISEGTKAVTKFSSSTN
P
H2B-1:
M/Z:1028.9874, Charge: 2+, ΔM: 1ppm, Ion score: 38
WT3_H2B_propionylated_2 #3593 RT: 57.39 AV: 1 NL: 2.78E3T: ITMS + c NSI d Full ms2 [email protected] [270.00-2000.00]
400 600 800 1000 1200 1400 1600 1800 2000m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Re
lativ
e A
bu
nd
an
ce
1011.57
954.181106.48789.17
1463.78 1679.73723.82 1373.60857.89666.41519.29 1593.65 1846.07378.06 1695.761207.33 1947.50
y8y10 y11
b3y13
y14y15
y16 y17y7,+2 y13,+2y17,+2
b3-H2O
b10
b11
[M-H2O+2H]2+
WT vs N15 WT (59.8:1)L
H
RT
Inte
nsi
ty
EZL2 vs N15 WT (25.2:1)
L
H
RT
Inte
nsi
ty
TXR1 vs N15 WT (12.9:1)
L
H
RT
Inte
nsi
ty
Final ratio
ΔEZL2/WT 0.42
ΔTXR1/WT 0.22
