Oxidaitive phosphorylation

5

TCA
-
ATP

proton motif force = pmf -
-'

' NADH, FADH2
4
. ' :.

?


.

ATP ,
. ATP
 4
:

NADH oxidoreductase/dehydrogenase .1
succinate dehydrogenase .2
(
cytochrome c oxidoreductase Q .3
cytochrome c oxidas .4
IV ,III ,I
'


) (donor ) (acceptor

:
.
.

.1 - .NAD+

NAD+ -

2e + 2 H + + NAD + H + + NADH

flavoproteins -FAD/FMN .2 ,flavin -

FMN + e + H + semiquinone _ int ermediate + e + H + FMNH 2

Ubiquinone = UQ = Coenzyme Q .3 ,
-

UQ + e + H + UQH * + e + H + UQH 2

.1 - , .

e + Fe 3+
Fe 2+
4 .
.
.
- .

.Iron-Sulfur proteins .2

, 4 ).(SH
, .
.
I.II -
 4 -
 III -
.NADH I
II-
.FADH2
UQ - . Fe-S
, I
)NADH + Q +5H+ NAD+ =QH2 + 4H(CYTOSOL

.II
FADH2 +Q FAD+ +QH2

I II
Fe-S Fe-S

FMNH2 FADH2
 Q-cycle III

Q
.C
UQ ,
-

UQH* = Semiquinone
 IV

4 C 4
IV ,
2 - .

IV .,

4 -
, 4
-.


4 :
.1 -
,
.
.2 .
 -
. -
Rotenon IUQ -
Antimycin A C1
,CN-,N3- CO
.

.
 ATP -
ATP
ET H + ATP H + = pH +
ATP

/FCCP

ATP
DCCD/
 ' 1
.1 Difference spectra - 4 ) (4-1-
- .
) (
.1-4
.
X +++ -
Y - - - -
, ) (1 ,2 ,3 ,4 ".
 ? ,
, .
),3 ,4 . -
(1 ,2 " " ,
.

:


/
. 3 2/1 4

: (- ) Y 4 1
" ' .
(- ) Y ) ' . ( ..
3 2 . 3 2 1 4 . 2

(+) X 4 1
2
3 ."

' . ?
.
 ' 2
, , ,MgCl2 , .ATP
- ) (Pi 10- (TMPD) .
.

__________________________________________________________________

)( )(
__________________________________________
O ________________________
2

91 15.0 Control . 1

76 15.1 . 2 1 )TMPD (0.3 mg

00 0.13 . 3 1 )Antimycin A (0.5 g

62 14.9 . 4 1 TMPD + Antimycin A

__________________________________________________________________

. , ATP - .
. , TMPD - )( )(.
. TMPD ATP .
. ) (P/O = P/2e ?
 . , ATP -

: . e
: ATP
ATP . ADP
ATP
. ADP
:
ATP
: Pi - . ATP
 __________________________________________________________________

)( )(
__________________________________________________________________

91 15.0 Control . 1

76 15.1 . 2 1 )TMPD (0.3 mg

00 0.13 . 3 1 )Antimycin A (0.5 g

62 14.9 . 4 1 TMPD + Antimycin A

__________________________________________________________________

. ) (P/O = P/2e .

P/O = P/2e = ATP

O2 +4e 2H2O" 15 . 2e 30 O2 O
.91/30=3/1" Pi 3 , = 3 ATP

TMPD ? TMPD ' , ' .

.
? 2.5/1. 76/15.1 ATP .

? 3
. 0
? . 0 . ATP ' _ (
.' .
TMPD? . 2/1 ' 2 ATP
__________________________________________________________________

) ( ) (
__________________________________________________________________
. ,
91 15.0 Control . 1 TMPD - )(
76 15.1 . 2 1 )TMPD (0.3 mg )(.
00 0.13 . 3 1 )Antimycin A (0.5 g . TMPD
62 14.9 . 4 1 TMPD + Antimycin A ATP .
__________________________________________________________________

TMPD ? TMPD e
-e" )
(
,
2.5" ATP ' )
' "
-' (3
? Antimicine ' QH2
cytochrome c oxidoreductase Q ) c1 , (3
0 ATP ADP . . 0
2 ? ,
-" . ATP 2/1
TMPD e . e"
/ . TMPD
= , , .
? TMPD
3 ' .
' TMPD
 ' 3
:
Succinate - 10mM
Pi - 10mM
ADP - 0.1mM
glucose - 10mM
Hexokinase + Mg2+ - 5mM

, - :

O2 consumed Pi disappeared Fe3(CN)63- reduced

)(moles )(moles )(moles
1 1.3 5.1 -
2 +KCN+ Fe3(CN)63- 0.0 2.5 5.1
+KCN+Fe3(CN)63- + 0.0 0.0 0.1
3
Antimycin A

. .
. . ,
.ATP
. A KCN - .ATP-
 . .

: , e " .FAD+
. FADH2
...
: ATP
ATP . ADP
ATP .
:
ATP
: ATP
= = =
.e
 . . ,
.ATP
. A KCN - -

. . e
: 5.1 ,2 2.6 2/1
? .
? , 4
.
2 ?

. ."
. ? e
- . )
) (4 ( - . ?
?

: ) (,
' 2 ,
." 5.1 2.55 = 2
2.5 . 1= 2.5 ,? .
+:
? ?
. ? 2) . 3
' Q ( C ? . 0?
. - ,
e ,
. .
) (
 .' C ) . (4
) ( ....
. ) ( Na+ -CN : C , 4
) . (
/ ." C ..
." . .
DNP . : C ) DNP
(
) "" .
( ..... C ."
DNP C . ....

C :3 , C
) 3 ( Q ' .4 3
) ( QH2 . 2&1

.....
 2 .1: ) NADH ( FADH2
.2 . pH
.... ? )(
)( ) (
. ) (
. ATP" :
NADH : FADH2
proton motif force = pmf :ATP . ATP"
.

. ) ,
NADH FADH2 , NADH FADH2
.
.
NADH FADH2 ... ...." .
 . :2 .
FADH2 , 2 . NADH
...

:
."
/ ...