'

12/ 1/1212 : : ,
' ' " : ' 205-1-9171

: . : ,
.
2 . .
.
. . ,
. 12' . '
' 3' . , .123
!
( 1 ) ( 03')
- ('-') . .
6.
6( .') .
,- galactosidase
.
. ,
. ( ,)WT
()
.)PEGylated( PEG ,
- galactosidase ( ).

Variant variation (M) Km Vmax )Kcat(S-1

number )(mmol\min
1 W.T 1350 0.01545 0.05
2 PEGylated 340 0.0165 0.047
3 E798L 1245 0.0126 0.042
4 W1000F 4500 3.73*103 0.007
5 E462H 20 2.05*103 0.12
, . kcat/KM
5<2<1<3<4

12/1/1212 ' , ' ' 1 11

 6( .')
.
,
? .
Glucagon (ligand) -> beta adrenergic receptor (receptor)-> Heterotrimeric Gs (G
>protein) -> Adenylate cyclase (effector) -> cAMP (second messanger) -> PKA -
PFK2 phosphorylation -> PFK2 inactiave, FBPase2 active -> F2,6BP
dephosphorylation -> no PFK stimulation -> glycolysis is off

. PFK
. ,
( )PEP carboxykinase, F1,6 biphosphatase
( Hexokinase, PFK, Pyruvate
.)kinase
6( .') ) phoshponoacetyl-L-aspartate (PALA
" .ATCase
PALA ? PALA
.ATCase

APAP ATCase
)1( : R ( )2
.
R

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 6( .') ( ) .

-
. .

Sphingomyelin

.

% .
.
.

6( .') ( )Size exclusion

pH=7 . . ,
( )cation exchange pH=7
.
. SP4 -SP1
.

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 ( ) . .
. ./
. SP1 ./
. SP4 ./

. SP2 ,SP1 SP3
./
. SP2 .SP4/
SP1 . ./

2 ( 22')
)(V0

P B
S .P
) )V0

.
.
-
.
)(mM

4( .') .

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4( .') , P B
( [S]c ). , . V0(A)=V0(B) ,[S]c
] [S]C>[S ) .V0(B)<V0(A , ] [S]C > [S ) .V0(A)> V0(B
P , kcat ,B ?
.
. A .
kcat A
(.)Vmax= kcat*E

4( .') S .0.3 mM
.

, 0.3 mM ,
B A

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 5( .') kcat/KM 4 kcat/KM .
, ? .

.
B .P B
. P
.

4( .') [S]c .

4( .') X
. ? ?X

. ,
, .
X , (
).

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 ( 0) ( 23')

( ) .
4'.

GLUT2 .1 GLUT2 .
:

. ER

. Km

. 6

.2 )PFK( Phosphofructokinase

PFK . citrate

ATP . .PFK

PMA . .PFK

P2,6BA . PFK

.3 ) (turnover number 100 sec-1

" .15 sec-1 ?
. " .
. " " "
.

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 .
" 6.6 .
. , ,
" .

.4 '-' ,
:
G __________ __________ ___________

. / / .
. / / .
. / / .
./GTPase / .
. /GTPase / .

.5 :
. .F-2,6-BP
. , ATP , .PFK
. ( )TIM ATP .
. F-1,6-BP .
. +
. +

12/1/1212 ' , ' ' 8 11

 4 ( 22')

. ' '
. ' '
11 ( )
11 ( ) .
( )Preference index 0 1 0
( 11) .
. ,
. ninaE

.ninaE A
. '

.ninaE ( )B ' 24

.ninaE

Wild type
No ninaE 1
No ninaE 2

12/1/1212 ' , ' ' 9 11

 ( 6') , A .
,ninaE ninaE
? , ninaE ? ,
? .

ninaE
11 26 . ninaE , .
ninaE
ninaE 26
32 .
.

6( .') ninaE
,
. .
, ninaE .
, ninaE .
.

. , .
.
, .
.

6( .') ninaE
, , ATP GTP
.
.
ATP .GTP .
.

ATP .
. GPCR
"
cAMP .

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 GTP .
G (
GTP .)GDP ( G )
.IRS GTP
.

6( .') P332 ninaE

.
. WT
.
.

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 :1

Standard Free Energy Changes

Reaction G0', kcal/mole

Phosphoenolpyruvate + H2O Pyruvate + Pi -12.8

3-Phosophoglyceroylphosphate + H2O 3-Phosphoglycerate + Pi -11.8

Acetyl phosphate + H2O Acetate + Pi -12.3

Phosphocreatine + H2O Creatine + Pi -12.3

Pyrophosphate + H2O 2 Pi -2.6

Acetyl-CoA + H2O Acetate + CoASH -7.2

ATP + H2O ADP + Pi -7.3

ATP + H2O AMP + Pyrophosphate -12.9

Glucose-1-phosphate + H2O Glucose + Pi -2.2

Fructose-1,6-diphosphate + H2O Fructose-6-phosphate + Pi -3.9

Glucose-6-phosphate + H2O Glucose + Pi -3.3

Glycerol-3-phosphate + H2O Glycerol + Pi -1.1

Glucose-6-phosphate Fructose-6-phosphate +2.2
2-Phosphoglycerate Phosphoenolpyruvate + H2O +2.2
3-Phosphoglycerate 2-Phosphoglycerate +1.1

Dihydroxyacetone phosphate Glyceraldhyde-3-phosphate +1.8

Fructose-1.6-diphosphate +2.7
Dihydroxyacetonephosphate + Glyceraldehyde-3-

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 phosphate

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 :3
)(Embden - Meyerhof

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