1bq1
From Proteopedia
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E. COLI THYMIDYLATE SYNTHASE MUTANT N177A IN COMPLEX WITH CB3717 AND 2'-DEOXYURIDINE 5'-MONOPHOSPHATE (DUMP)
Overview
Mutation of thymidylate synthase N229(177) to alanine results in an, essentially inactive enzyme, yet it leads to formation of a stable ternary, complex. The kinetics of N229(177)A show that kcat for Escherichia coli is, reduced by 200-fold while the Km for dUMP is increased 200-fold and the Km, for folate increased by tenfold versus the wild-type enzyme. The crystal, structures of N229(177)A in complex with dUMP and CB3717, and in complex, with dUMP alone are determined at 2.4 A, and 2.5 A resolution. These, structures identify the covalently bound ternary complex and show how, N229(177)A traps an intermediate, and so becomes inactive in a later step, of the reaction. Since the smaller alanine side-chain at N229(177)A does, not directly sterically impair binding of ligands, the structures, implicate, and place quantitative limits on the involvement of the, structured water network in the active site of thymidylate synthase in, both catalysis and in determining the binding affinity for dUMP (in, contrast, the N229(177)V mutation in Lactobacillus casei has minimal, effect on activity).
About this Structure
1BQ1 is a Single protein structure of sequence from Escherichia coli with UMP and CB3 as ligands. Active as Thymidylate synthase, with EC number 2.1.1.45 Full crystallographic information is available from OCA.
Reference
Inactivity of N229A thymidylate synthase due to water-mediated effects: isolating a late stage in methyl transfer., Reyes CL, Sage CR, Rutenber EE, Nissen RM, Finer-Moore JS, Stroud RM, J Mol Biol. 1998 Dec 4;284(3):699-712. PMID:9826509
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