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Literature summary for 3.5.1.2 extracted from

  • DeLaBarre, B.; Gross, S.; Fang, C.; Gao, Y.; Jha, A.; Jiang, F.; Song J, J.; Wei, W.; Hurov, J.B.
    Full-length human glutaminase in complex with an allosteric inhibitor (2011), Biochemistry, 50, 10764-10770.
    View publication on PubMed
Search for more literature for 3.5.1.2

Cloned(Commentary)

Cloned (Comment) Organism
expression of GAC in Spodoptera frugiperda Sf9 cells Homo sapiens

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant GAC free and in complex with inhibitor bis-2-(5-phenylacetimido-1,2,4,thiadiazol-2-yl)ethyl sulfide, hanging drop vapor diffusion, mixing of protein in 25 mM HEPES (pH 7.5), 200 mM NaCl, 5% glycerol, and 5 mM beta-ME, with reservoir solution containing 0.3 M magnesium chloride, 0.1 M Tris, pH 8.5, and 12% w/v PEG 4000, X-ray diffraction structure determination and analysis at 2.55 and 2.3 A resolution, respectively Homo sapiens

Protein Variants

Protein Variants Comment Organism
F322S/F318Y site-directed mutagenesis, mutant of GLS1 splice variant GAC, the mutant is not inhibited by bis-2-(5-phenylacetimido-1,2,4,thiadiazol-2-yl)ethyl sulfide in comtrast to the wild-type enzyme Homo sapiens
Y394L site-directed mutagenesis, mutant of GLS1 splice variant GAC, the mutant is not inhibited by bis-2-(5-phenylacetimido-1,2,4,thiadiazol-2-yl)ethyl sulfide in comtrast to the wild-type enzyme Homo sapiens

Inhibitors

Inhibitors Comment Organism Structure
bis-2-(5-phenylacetimido-1,2,4,thiadiazol-2-yl)ethyl sulfide inhibits specifically GLS1 and its splice variant GAC, two inhibitor molecules bind at an interface region of the GAC tetramer in a manner that appears to lock the GAC tetramer into a nonproductive conformation, binding structure and mechanism of glutaminase inhibition, overview; inhibits specifically GLS1 splice variant GAC, two inhibitor molecules bind at an interface region of the GAC tetramer in a manner that appears to lock the GAC tetramer into a nonproductive conformation, binding structure and mechanism of glutaminase inhibition, overview Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.4
-
 L-glutamine pH 8.5, temperature not specified in the publication, GAC mutant Y394L Homo sapiens
1.4
-
 L-glutamine pH 8.5, temperature not specified in the publication, wild-type GAC Homo sapiens
1.9
-
 L-glutamine pH 8.5, temperature not specified in the publication, isozyme GLS1 Homo sapiens
2.5
-
 L-glutamine pH 8.5, temperature not specified in the publication, GAC mutant F322S/F318Y Homo sapiens
4
-
 L-glutamine pH 8.5, temperature not specified in the publication, isozyme GLS2 Homo sapiens

Localization

Localization Comment Organism GeneOntology No. Textmining
mitochondrion
-
 Homo sapiens 5739
-

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-glutamine + H2O Homo sapiens
-
 L-glutamate + NH3
-
 ?

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
 isozyme GLS2
-
Homo sapiens O94925 GAC; GAC, a splice variant of isozyme GLS1
-
Homo sapiens O94925 GLS1; isozyme GLS1
-

Purification (Commentary)

Purification (Comment) Organism
recombinnat GAC from Spodoptera frugiperda Sf9 cells by nickel affinity chromatography Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-glutamine + H2O
-
 Homo sapiens L-glutamate + NH3
-
 ?
L-glutamine + H2O the glutamate bindig pocket of the enzyme involves Glu381 and Tyr249, structure overview Homo sapiens L-glutamate + NH3
-
 ?

Subunits

Subunits Comment Organism
tetramer GLS1 splice variant GAC Homo sapiens

Synonyms

Synonyms Comment Organism
GAC splice variant of GLS1 Homo sapiens
GLS1
-
 Homo sapiens
GLS2
-
 Homo sapiens
glutaminase 1
-
 Homo sapiens
glutaminase 2
-
 Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.5
-
 L-glutamine pH 8.5, temperature not specified in the publication, isozyme GLS2 Homo sapiens
10
-
 L-glutamine pH 8.5, temperature not specified in the publication, isozyme GLS1 Homo sapiens
20
-
 L-glutamine pH 8.5, temperature not specified in the publication, GAC mutant Y394L Homo sapiens
22
-
 L-glutamine pH 8.5, temperature not specified in the publication, wild-type GAC Homo sapiens
40
-
 L-glutamine pH 8.5, temperature not specified in the publication, GAC mutant F322S/F318Y Homo sapiens

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.00006
-
 pH 8.5, temperature not specified in the publication, isozyme GLS1 Homo sapiens bis-2-(5-phenylacetimido-1,2,4,thiadiazol-2-yl)ethyl sulfide
0.00008
-
 pH 8.5, temperature not specified in the publication, wild-type GAC Homo sapiens bis-2-(5-phenylacetimido-1,2,4,thiadiazol-2-yl)ethyl sulfide
0.088
-
 pH 8.5, temperature not specified in the publication, isozyme GLS2 Homo sapiens bis-2-(5-phenylacetimido-1,2,4,thiadiazol-2-yl)ethyl sulfide
0.1
-
 pH 8.5, temperature not specified in the publication, GAC mutant F322S/F318Y and GAC mutant Y394L Homo sapiens bis-2-(5-phenylacetimido-1,2,4,thiadiazol-2-yl)ethyl sulfide

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.63
-
 L-glutamine pH 8.5, temperature not specified in the publication, isozyme GLS2 Homo sapiens
5.3
-
 L-glutamine pH 8.5, temperature not specified in the publication, isozyme GLS1 Homo sapiens
16
-
 L-glutamine pH 8.5, temperature not specified in the publication, wild-type GAC and GAC mutant F322S/F318Y Homo sapiens
47
-
 L-glutamine pH 8.5, temperature not specified in the publication, GAC mutant Y394L Homo sapiens