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PDBsum entry 2w1v
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References listed in PDB file
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Key reference
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Title
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Functional proteomic and structural insights into molecular recognition in the nitrilase family enzymes.
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Authors
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K.T.Barglow,
K.S.Saikatendu,
M.H.Bracey,
R.Huey,
G.M.Morris,
A.J.Olson,
R.C.Stevens,
B.F.Cravatt.
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Ref.
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Biochemistry, 2008,
47,
13514-13523.
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PubMed id
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Abstract
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Nitrilases are a large and diverse family of nonpeptidic C-N hydrolases. The
mammalian genome encodes eight nitrilase enzymes, several of which remain poorly
characterized. Prominent among these are nitrilase-1 (Nit1) and nitrilase-2
(Nit2), which, despite having been shown to exert effects on cell growth and
possibly serving as tumor suppressor genes, are without known substrates or
selective inhibitors. In previous studies, we identified several nitrilases,
including Nit1 and Nit2, as targets for dipeptide-chloroacetamide activity-based
proteomics probes. Here, we have used these probes, in combination with
high-resolution crystallography and molecular modeling, to systematically map
the active site of Nit2 and identify residues involved in molecular recognition.
We report the 1.4 A crystal structure of mouse Nit2 and use this structure to
identify residues that discriminate probe labeling between the Nit1 and Nit2
enzymes. Interestingly, some of these residues are conserved across all
vertebrate Nit2 enzymes and, conversely, not found in any vertebrate Nit1
enzymes, suggesting that they are key discriminators of molecular recognition
between these otherwise highly homologous enzymes. Our findings thus point to a
limited set of active site residues that establish distinct patterns of
molecular recognition among nitrilases and provide chemical probes to
selectively perturb the function of these enzymes in biological systems.
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