HEMOGLOBIN, 11(1), 6 1 - 6 2 ( 1 9 8 7 )
BRIEF REPORT
HEMOGLOBIN SWAN RIVER [a6(A4)ASP+GLY]
Hemoglobin Swan R i v e r (1) was de tec ted i n a 24-year-o ld female Caucasian. The v a r i a n t appeared as a band w i t h a m o b i l i t y between Hbs F and S on c e l l u l o s e a c e t a t e e l e c t r o p h o r e s i s (pH 8.6). A v a r i - a n t Hb A2 band c a t h o d i c t o normal Hb Ap was a l s o observed, i n d i - c a t i n g t h e presence o f an a c h a i n v a r i a n t . C i t r a t e agar e lec t ropho- r e s i s a t pH 6.0 revea led a band between Hbs A and S , and g l o b i n cha in a n a l y s i s i n a c i d and a l k a l i n e b u f f e r s showed an abnormal c h a i n m i g r a t i n g between t h e aA and aG P h i l a d e l p h i a chains (2 ) . The i s o - propanol t e s t ( 3 ) f o r uns tab le hemoglobins was p o s i t i v e as was t h e BCB t e s t ( 4 ) f o r i n c l u s i o n bodies. Q u a n t i t a t i o n o f a hemolysate on Mono Q a n i o n i c r e s i n (Pharmacia) r e s u l t e d i n 88.2% Hb A, 9.8% Hb Swan R ive r , and 2.0% Hb A2 (5 ) . The percentage o f Hb F was 0.5%. We were unable t o o b t a i n any hemato log ic da ta on t h i s p a t i e n t o r on h e r two c h i l d r e n who a l s o have t h e v a r i a n t .
The v a r i a n t hemoglobin was p a r t i a l l y r e s o l v e d by chromatography on DEAE-Sephadex ( 6 ) as a shoulder peak e l u t i n g ahead o f Hb A. Separat ions o f t h i s f r a c t i o n on CM-cel lu lose ( 7 ) gave a ma jo r peak o f s lower e l u t i n g v a r i a n t a c h a i n c l e a r l y r e s o l v e d f rom a m ino r peak c o n t a i n i n g t h e cJ\ chain. The v a r i a n t c h a i n was aminoe thy la ted (8) , d iges ted w i t h t r y p s i n (9), and f i n g e r p r i n t e d (10) . The p e p t i d e map i n d i c a t e d a change i n t h e p o s i t i o n o f Tp-1. T h i s p e p t i d e t h a t n o r m a l l y i s s i t u a t e d above Tp-9 was now l o c a t e d between Tp-6 and Tp-3 (11). These r e s u l t s r e v e a l e d t h a t abnormal Tp-1 was more hydrophobic and l e s s a c i d i c than t h e normal p e p t i d e was exc i sed f rom t h e map, e l u t e d and hydro lyzed. The amino a c i d compos i t i on o f t h i s pep t ide : Asp 0.0(1), Ser l . O ( l ) , Pro 0.93(1), Gly 1.0(0), A l a 0.97(1), Val l.l(l), Leu 0.7(1) and Lys 1.3(1) t h a t shows t h e s u b s t i t u t i o n o f a r e s i d u e o f g l y c i n e f o r a s p a r t i c a c i d would account f o r i t s a l t e r e d m o b i l i t y . The s u b s t i t u t i o n o f g l y c i n e f o r t h e s i n g l e r e s i d u e o f a s p a r t i c a c i d a t p o s i t i o n 6 i n Tp-1 was con f i rmed b y sequence a n a l y s i s o f t h e whole c h a i n on a Beckman 890C sequencer.
F i v e o t h e r v a r i a n t s have been desc r ibed a t t h i s p o s i t i o n : Hb Sawara (Asp-Ala) (12) , Hb Dun ( A s p A s n ) (13), Hb Ferndown ( A s p V a l ) (14) , Hb Woodvi l le (Asp+Tyr) (151, and Hb Boyle H e i g h t s (Asp-4) ( 6 ) . A l l show inc reased oxygen a f f i n i t y t h a t has been a t t r i b u t e d t o t h e l o s s o f t h e c a r b o x y l a t e which forms hydrogen bonds
Received : Mahch 3, 1986; Accepted : O&ube,t 24 , 1986 .
61
Copyright @ 1987 by Marcel Dekker, Inc.
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62 MOO-PENN ET AL.
with the OH of Ser 3(Al)a and Ser 124(H7)a. Loss of these bonds may loosen the structure as a whole (17). The cooperativity and Bohr effect were unchanged for those variants where these properties were reported.
We wish to thank S. Holland and H. Brown for technical assist- ance.
Division of Host Factors Center for Infectious Diseases Centers for Disease Control Atlanta, GA 30333
Chemical Sciences Division Texas Department o f Health Austin, TX 78756
W. F. Moo-Penn D.L. Jue
M.H. Johnson
B. L. Therrell
1. 2. 3. 4.
5. 6. 7.
8.
9. 10. 11.
12.
13.
14.
15.
16.
17.
REFERENCES
Wrightstone, R.N., Hemoglobin, 9:227, 1985. Schneider, R.G., Crit. Rev. Clin. Lab. Sci., 9:203, 1978. Carrell, R.W. and Kay, R., Br. J. Haematol., 23:615, 1972. Schneider, R.G., Takeda, I., Gustavson, L.P., and Alprin, J.B., Nature (New Biol.), 235:88, 1972. Moo-Penn, W.F. and Jue, D.L., J. Chromatogr., 318:325, 1985. Huisman, T.H.J. and Dozy, A.M., J. Chromatogr., 19:160, 1965. Clegg, J.B., Naughton, M.A., and Weatherall, D.J., J. Mol. Biol., 19:91, 1966. Raftery, M.A. and Cole, R.D., Biochem. Biophys. Res. Commun., 10:467, 1963. Smyth, D.G., Methods Enzymol., 11:214, 1967. Bennett, J.C., Methods Enzymol., 11:330, 1967. Winter, W.P. and Rucknagel, D.L., in The Detection of Hemo- jlobinopathies, edited by R.M. Schmidt, T.H.J. Huisman, and H. Lehmann, page 51, CRC Press, Boca Raton, FL, 1974. Sasaki, J., Imamura, T., Sumida, I., Yanase, T., and Ohya, M., Biochim. Biophys. Acta, 495:183, 1977. Jue, D. L., Johnson, M. H. , Patchen, L . C . , and Moo-Penn, W. F., Hemoglobin, 3:137, 1979. Lee-Potter, J.P., Deacon-Smith, R.A., Lehmann, H., and Robb, L., FEBS Lett., 126:117, 1981. Como, P. F, Barber, S., Sage, R.E., Trent, R. J., and Kronenberg, H., Hemoglobin, 10:135, 1986. Johnson, C.S., Schroeder, W.A., Shelton, J.B., and Shelton, J.R., Hemoalobin, 7:125, 1983. Fermi, G . and Perutz, M.F., Haemoglobin and Myoglobin, Claren- don Press, Oxford, 1981.
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