2oip

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Revision as of 11:49, 23 January 2008

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spinqualitylabelsall models 2oip, resolution 2.80Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image

Crystal Structure of the S290G Active Site Mutant of TS-DHFR from Cryptosporidium hominis

Overview

Cryptosporidium hominis TS-DHFR exhibits an unusually high rate of, catalysis at the TS domain, at least 10-fold greater than those of other, TS enzymes. Using site-directed mutagenesis, we have mutated residues, Ala287 and Ser290 in the folate-binding helix to phenylalanine and, glycine, respectively, the corresponding residues in human and most other, TS enzymes. Our results show that the mutant A287F, the mutant S290G, and, the double mutant all have reduced affinities for methylene, tetrahydrofolate and reduced rates of reaction at the TS domain., Interestingly, the S290G mutant enzyme had the lowest TS activity, with a, catalytic efficiency approximately 200-fold lower than that of the wild, type (WT). The rate of conformational change of the S290G mutant is, approximately 80 times slower than that of WT, resulting in a change in, the rate-limiting step from hydride transfer to covalent ternary complex, formation. We have determined the crystal structure of ligand-bound S290G, mutant enzyme, which shows that the primary effect of the mutation is an, increase in the distance between the TS ligands. The kinetic and crystal, structure data presented here provide the first evidence explaining the, unusually fast TS rate in C. hominis.

About this Structure

2OIP is a Single protein structure of sequence from Cryptosporidium hominis with , , and as ligands. Full crystallographic information is available from OCA.

Reference

Nonconserved Residues Ala287 and Ser290 of the Cryptosporidium hominis Thymidylate Synthase Domain Facilitate Its Rapid Rate of Catalysis(,)., Doan LT, Martucci WE, Vargo MA, Atreya CE, Anderson KS, Biochemistry. 2007 Jul 17;46(28):8379-91. Epub 2007 Jun 20. PMID:17580969

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