1.

2.

3. DNA

7o

210

1. O

25 o () . (NOS, Nitric Oxide Synthases) L-. , . .

1, . , (sGC, soluble Guanylyl Cyclase). , sGC cGMP GTP. , cGMP , .. (PKG, PKA), (PDE) (CNG).

1, , DNA. , (nitrosonium ion, NO+) (Cys) (Cys-NO). , (peroxynitrite, ONOO -) (Tyr-NO2). , DNA.

, (NOS, Nitric Oxide Synthase). NOS , L- 2 , L- .

7.1 , S.

211

, NADPH , FMN, FAD, , , Ca2+/ Zn2+ .

7.2 NOS, .

3 NOS: NOS I, NOS II NOS III. NOS I NOS (nNOS), , ,

. NOS II, ,

NOS (iNOS), , Ca2+/ .

, NOS III, , , , NOS (eNOS).

, NOS o, . NOS, NOS knockout .

NOS I knockout , ( ) .

NOS II knockout . NOS II LPS, Gram , NOS .

NOS III knockout , , NOS III . , NOS II , NOS III .

212

: .

2 , 1 , 2,5. , .

, , . . , 1,4 37C, , , , ( ) .

>1

213

2.

, , , . 3 : , (sGC). sGC , , , , , .

Fe-S : SoxR, HIF-1.

1. SoxR Escherichia coli , stress. 2Fe-2S SoxR dinitrosyl-iron-dithiol , SoxR .

2. . , 4Fe-4S

() Krebs, .

, , , Fe 4Fe-4S. 4Fe-4S IRP (Iron-Regulatory Protein). IRP IREs (Iron-Responsive Elements) 5- mRNA 3- mRNA . IRP mRNA , mRNA . , . IRP, Fe .

3. O HIF-1 (Hypoxia Inducible Factor) (.. VEGR: Vascular Endothelial Growth Factor). HIF-1 : HIF-1 ARNT. , HIF-1 VEGR VEGR.

1. -1 , (CO) . mRNA -1 -1 . -1 NOS, NOS . , , NOS

214

mRNA -1 . , , feedback .

2. c . , . c , , . , c .

3. (H2O2) . , , .

(sGC) sGC . :

GTP c GMP + PPi , . 2 (1, 2, 1, 2). 3 :

1. NH2 . 2. . 3. COOH . sGC

: NH2 2 , COOH 2 .

s GC 3 :

1. , . Fe2+ . His105 1 .

2. Fe2+ -Fe2+ Fe2+ . sGC 4 .

3. , Fe2+ His105 1 , Fe2+ . 100 , 400 ( 7.3).

7.3 E sGC .

215

sGC

YC-1: H . NO , .

ODQ: A Km Vmax . NO Fe2+ Fe3+ .

BAY 41-2272: sGC, . cGMP

1. o cGMP , . cGMP , cGMP (PKG). 2 PKGs: PKG I, PKG II. PKG I. 7.1.

7.1 PKG I.

, PKG I ( 7.4).

, . . .

PKG I

IP3 Ca2+, .

Ca2+-ATP .

Ca2+, .

2 .

cAMP Response Element Binding Protein

Fos

.

, .

216

7.4 . GPCR , C, 3. 3 Ca2+, . . , . , , cGMP. cGMP PKG, 2 ( Ca2+ , MLCP, ) , .

2. cGMP cAMP (PKA). 50 PKA cAMP.

3. cGMP PKG I, cGMP (CNG). 5 CNG. CNG 1 3 , . , CNG 3, , .

4. (PDEs) cGMP. cAMP cGMP 5-AMP 5-GMP . 5 PDEs. , cGMP (PDE 5), . PDE 5 VIAGRA. Viagra, PDE 5, cGMP PKG I, .

217

3.

micromole. - 2. 2 . . , 2 . , nanomole, o 70 , micromole, .. , 2 0,2 .

2 , 2 23 (dinitrogen trioxide). 23 + (nitrosonium ion) 2- (nitrite ion). + , .

, 2 +, , , +.

2NO + O2 2NO2 NO + NO2 N2O3 SNO NO+ NO2- R-SH

7. 5 . - 2. 2

, 2 23 (dinitrogen trioxide). 23 + (nitrosonium ion) 2- (nitrite ion). + , .

. GTP p21 (RAS)

GTP p21 ( Ras). Ras , . GDP GTP. Ras 5 , 118 . Ras , (mitogen activated protein kinases) 3- (phosphatidylinsitol-3 kinase). 118 .

218

PC12 Ras C118S Ras, NGF (nerve growth factor). NGF . , Ras - 3-, .

. N 3

3 (caspase 3), . , .

Fas, 3, , . 3 . Jurkat T 3 . NOS L-NAME (N-nitro-L-arginine methyl ester) , 3 , .

- 3 3 . , .

. , NMDA (N-Methyl-D-

Aspartate) (ryanodine), in vivo. O NMDA, 5 , , ( ). NMDA . :

, Mg2+. non-NMDA , a+ . Mg2+ NMDA , Ca2+ Na+. NOS , NO.

, 399 NR2A DA. A . , , .

219

7.6 NMDA

,

Ca2+ . - - , . - , Ca2+ . Ca2+ , .

84 , 12 , .

220

7.7

. H . NF-kB p50 p65 . 1-kB. , 1-kB 1-kB, 1-kB NF-kB

, DNA .

p50 F-kB.

7.8 F-kB

221

IRPs, . ce 1 8 4 . ce 1 CUP, . , , CUP Ace 1. , Ace 1 .

7.9 ce 1.

, ce 1 CUP, .

,

. CpG DNA . DNA DNA.

. (ONOO-, peroxynitrite), . mole NO mole (superoxide radical, 2-), ONOO-, (superoxide dismutase, SOD). H / , / , 22, .

222

80% pKa=6,8 20% NO + O2 - CIS (ONOO-) TRANS (ONOOH) pKa=7,6 Tyr-NO2 Tyr OH + NO2

NO2 H2O Tyrosine

7.10 .

- To (-) . - , - (peroxynitrous acid). H pka 6,8 p 7,4, 80% - Henderson-Hasselbach. , - cis - trans . trans , cis - . . trans 2 - (hydroxyl radical). , Fe-S, 8- .

SOD Cu/Zn. . , 2, 3- (yr-NO2). , . in vivo in vitro.

7.2 In vivo

Prostacyclin Synthase(185-187)

Mn superoxide dismutase(188)

SERCA2A(189)

Neurofilament L(171)

Prostaglandin H Synthase(190-192)

Ribonucleotide Reductase(193-196)

Succinyl CoA:3-oxo acid CoA transferase(197)

(de-nitrase).

223

- .

, , , Alzheimer, Parkinson, .

. o NOS II. H NOS II SOD Cu/Zn - . o NOS II. NOS II LDL.

H ALS. To 10% ALS 20% SOD Cu/Zn. M SOD, . - L.

DNA To NO+ , DNA .

Hanafy, K.A. Krumenacker J.S. and F. Murad, NO, nitrotyrosine, and cyclic GMP in signal transduction, Med. Sci. Monit, 2001, 7, 801-819

Kleppisch T, Feil R., cGMP signalling in the mammalian brain: role in synaptic plasticity and behavior, Handb Exp Pharmacol. 2009 (191):549-79.

224