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Literature summary for 2.7.2.11 extracted from

  • Sekine, T.; Kawaguchi, A.; Hamano, Y.; Takagi, H.
    Desensitization of feedback inhibition of the Saccharomyces cerevisiae gamma-glutamyl kinase enhances proline accumulation and freezing tolerance (2007), Appl. Environ. Microbiol., 73, 4011-4019.
    View publication on PubMedView publication on EuropePMC

Activating Compound

Activating Compound Comment Organism Structure
additional information GK activity requires gamma-glutamyl phosphate reductase Saccharomyces cerevisiae

Application

Application Comment Organism
additional information GK is the rate-limiting enzyme in the proline biosynthesis Saccharomyces cerevisiae

Cloned(Commentary)

Cloned (Comment) Organism
expression of PRO1 in Escherichia coli M15(pREP4) Saccharomyces cerevisiae

Protein Variants

Protein Variants Comment Organism
D154N is 64fold less sensitive to feedback inhibition than wild-type GK, leading to proline accumulation Saccharomyces cerevisiae
E149K very insensitive to feedback inhibition, shows prominent increase in the proline content, freeze tolerance like the D154N mutant Saccharomyces cerevisiae
I150T very insensitive to feedback inhibition, shows prominent increase in the proline content, is more tolerant to freezing stress than cells expressing the D154N mutant Saccharomyces cerevisiae
N142D/I166V very insensitive to feedback inhibition, shows prominent increase in the proline content, is more tolerant to freezing stress than cells expressing the D154N mutant Saccharomyces cerevisiae
S146P shows prominent increase in the proline content Saccharomyces cerevisiae

Inhibitors

Inhibitors Comment Organism Structure
L-proline GK activity is subject to feedback inhibition by proline Saccharomyces cerevisiae
additional information no significant differences in growth between mutant yeast cells on SD agar medium with heat shock (50°C, 3 to 5 h), ethanol (16 to20%), and sorbitol (2.0 to 2.5 M) stresses, although the proline-accumulating cells are more resistant to these stresses than cells expressing wild-type GK Saccharomyces cerevisiae

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
47000
-
gel filtration Saccharomyces cerevisiae

Organism

Organism UniProt Comment Textmining
Saccharomyces cerevisiae P32264 strain INVDput1pro1
-

Purification (Commentary)

Purification (Comment) Organism
by Ni2+ column and gel filtration Saccharomyces cerevisiae

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2
-
mutant N142D/I166V Saccharomyces cerevisiae
4.2
-
mutant E149K Saccharomyces cerevisiae
14.7
-
wild-type Saccharomyces cerevisiae
26.3
-
mutant D154N Saccharomyces cerevisiae
29
-
mutant I150T Saccharomyces cerevisiae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + L-glutamate
-
Saccharomyces cerevisiae ADP + L-glutamate 5-phosphate
-
?

Synonyms

Synonyms Comment Organism
gamma-glutamyl kinase
-
Saccharomyces cerevisiae
GK
-
Saccharomyces cerevisiae

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.5
-
wild-type Saccharomyces cerevisiae L-proline
32
-
mutant D154N Saccharomyces cerevisiae L-proline
50
-
mutant I150T Saccharomyces cerevisiae L-proline
54
-
mutant N142D/I166V Saccharomyces cerevisiae L-proline
840
-
mutant E149K Saccharomyces cerevisiae L-proline