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,
, ,,
, ,,
,
NADPH-
NADPH-,
, .
.
,
, (,
( ,,
,
A,
A,C,
C,E,
E,,
,N-
N-
,),
, ),
x
x
-
-
AO
AO
( Cu/Zn Mn):
2- + 2- + 2+ 22 + 2
( ):
222 22 + 2
( ):
2GSH + 22 GSSG + 22O
( FAD):
GSSG + 2 2GSH + 2
(. )
(. )
(. )
N-
,
-
/
0,2
1,6 ()
/
1,95
Knock-out
Glu-peroxidase -/-
202 202
+ BSO (
-) + 3-
+/- 3- ( +/- S
)
GSH
-L-glutamyl-L-cysteinylglycine
1 - 10
~ 1 - 5 ,
(
)
,
GSH
(
GSH )
GluCys
(GSSG),
(NADPH), GSH/GSSG
1000/1
( 40-50% ,
, )
GSH
-
(Saer et al.,1990; Winterbourn, 1994);
,
(Chance et al., 1979)
(Cotdrave and
Gudes, 1997)
,
, (Meister
and Anderson, 1983; Cooper, 1997)
(Pool et al,. 1995)
(Chibelli et al., 1998; Hall, 1999)
NEW! (
; SH
NMDA ) (Janaky et al., 1999)
NEW!
(Paolicchi et al., 2002)
in vitro
- GSH +GSH
CDNB (chloro-2,4- et-GSH (
dinitrobenzene)
+
DEM (diethyl maleimide)
GSH )
,
-S-
DEM 1 mM, 1h
GSH CDNB - 1 mM, 30 min
( ) Et-GSH 5 mM, 30 min
CMFD (chloromethyl fluorescein ) N=5
GSH
CDNB ,
,
Na,K-, , ,
200
150
DHRfluorescencerelatedtocontrol, %
100
50
+CDNB 70
60
protein
50
40
ATP, nM/mg
0 30
Control CDNB DEM et-GSH 20
10
123(DHR) dihydrorhodamine, 0
0 5 10 15 20 25
rhodamine 123,
Incubation time, min
()
, , 1932 .,
De Vigneaud
- ,
, [Welch G.,
1998].
(1%),
(70%).
,
[ .., .., 2002].
,
15 /.
1530 / ,
30 100 / ,
100 500 / [Welch G., Loscalo J., 1998; Warren C., 2002].
-
-
-
( (
)
) 1975 . McCully
(
)
-
( )
( -
)
,
---------------------
In vitro
in vitro
:
PI ( ) ex=485 , em=610 (
)
130
125
120
115
DCF, %
110
105
100
95
MK-801 90 130
85 125
80 120
115
DCF, %
110
105
100
95
AIDA 90
85
80
I
MSOP III
In vivo
Experimental protocol
Behavioral test
1 6 14 2
()
23,3 0,4
( 4
10,01 /
2
(6 2)
() ) 71 18,9 0,5
B
3 +
0,10,01 /
( + 6 11 2 22,9 0,9
)
Content of HC in the blood of rats
under methionine
over-loading
NMDA,
HCA HC
500 30
1. , ,
.
2.
NMDA, HC HCA .
HC HCA
-NMDA
AIDA -
mGlu 1
MSOP
mGlu3
R .G. M. Morris et al. 1982. Nature, 297, 681-683.
,
:
1)
( );
2) ( );
3) ( /c);
4)
,
( %
);
5)
( )
( ),
2 - 4
2 - 4 .
, .
:
4 -5
1 2 3
( () () ()
)
20 7 140 18 45 6
,
, 52 20 5 8 2
, 0,24 0,02 0,18 0 02 0,25 0,04
/c
20 7 75 35 5
, %
( Cu/Zn Mn):
2- + 2- + 2+ 22 + 2
( ):
222 22 + 2
( ):
2GSH + 22 GSSG + 22O
( FAD):
GSSG + 2 2GSH + 2
(. )
(. )
(. )
N-
,
GSH
GSH
1) GSH+ Cu2+/Fe3+ Cu+/Fe2+ + GS + H+
2 - 2 - -
22 - - - + 2+ 2
+ NO +2+
+ 2 - + Fe2+
ONOO- 22 + 2
2 + - + - + - + Fe3+
( )
( -)
DCFH2 DCF
-
DCF
,
100 M
PI versus DCF coordinates
PI
ROS signal
300
Fluorescence, % to Control
3-HPG
250
NMDA
200
ACPD
150
100
50
0
0 100 250 500
Concentration of Ligands, mkM
Effect of
Effect of Glutamate
Glutamate Ligands
Ligands
on Na/K-ATPase
on Na/K-ATPase
160
3-HPG
140
Na/K-ATPase, % to Control
120
100
NMDA
80
60
ACPD
40
20
0
0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 1.0
Ligand Concentration, mM
SH-SY5Y
1
(2+3)
D Aktivated PkB, %
-
14-3-3
Kulikov et al,
Biochemistry, submitted
Na-
+D-AP5
NMDA-
MAPK
NMDA-
(* p<0.05 )
2+3 1
NMDA K+
K+
Ca2+ MAP
MAPK
K
IP3K
Ca2+ NADPH
Bcl2
kC
kC 14-3-3 +
k
k p53P -
.
OH
Fe2+
.-
02 202 Glu-SH NAD
20 GSSG NADH2
.
OH
Fe2+
.-
02 202 Glu-SH NAD
20 GSSG NADH2
GS-SG NAD
*
GluSH
-Tocoferyl*
NADH2
*
-Tocoferol Ascorbat
Asc*