Abstract
Glutaminase (GLS1/2) catalyzes the conversion of L-glutamine to L-glutamate and ammonia. The level of a splice variant of GLS1 (GAC) is elevated in certain cancers, and GAC is specifically inhibited by bis-2-(5-phenylacetimido-1,2,4,thiadiazol-2-yl)ethyl sulfide (BPTES). We report here the first full-length crystal structure of GAC in the presence and absence of BPTES molecules. Two BPTES molecules bind at an interface region of the GAC tetramer in a manner that appears to lock the GAC tetramer into a nonproductive conformation. The importance of these loops with regard to overall enzymatic activity of the tetramer was revealed by a series of GAC point mutants designed to create a BPTES resistant GAC.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Allosteric Site*
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Amino Acid Sequence
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Amino Acid Substitution
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Biocatalysis
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Databases, Protein
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Dimerization
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Enzyme Inhibitors / chemistry*
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Enzyme Inhibitors / metabolism*
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Glutaminase / antagonists & inhibitors*
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Glutaminase / chemistry*
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Glutaminase / genetics
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Glutaminase / metabolism
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Humans
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Isoenzymes / antagonists & inhibitors
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Isoenzymes / chemistry
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Isoenzymes / genetics
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Isoenzymes / metabolism
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Kinetics
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Models, Molecular
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Molecular Sequence Data
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Mutagenesis, Site-Directed
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Mutant Proteins / antagonists & inhibitors
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Mutant Proteins / chemistry
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Mutant Proteins / metabolism
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Point Mutation
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Protein Conformation
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Recombinant Proteins / antagonists & inhibitors
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Recombinant Proteins / chemistry
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Recombinant Proteins / metabolism
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Sequence Alignment
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Sulfides / chemistry
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Sulfides / metabolism
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Thiadiazoles / chemistry
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Thiadiazoles / metabolism
Substances
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Enzyme Inhibitors
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Isoenzymes
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Mutant Proteins
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Recombinant Proteins
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Sulfides
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Thiadiazoles
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bis-2-(5-phenylacetamido-1,2,4-thiadiazol-2-yl)ethyl sulfide
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Glutaminase