1tmh
MODULAR MUTAGENESIS OF A TIM-BARREL ENZYME: THE CRYSTAL STRUCTURE OF A CHIMERIC E. COLI TIM HAVING THE EIGHTH (BETA-ALPHA)-UNIT REPLACED BY THE EQUIVALENT UNIT OF CHICKEN TIM
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OverviewOverview
The crystal structure of a hybrid Escherichia coli triosephosphate, isomerase (TIM) has been determined at 2.8 A resolution. The hybrid TIM, (ETIM8CHI) was constructed by replacing the eighth beta alpha-unit of E., coli TIM with the equivalent unit of chicken TIM. This replacement, involves 10 sequence changes. One of the changes concerns the mutation of, a buried alanine (Ala232 in strand 8) into a phenylalanine. The ETIM8CHI, structure shows that the A232F sequence change can be incorporated by a, side-chain rotation of Phe224 (in helix 7). No cavities or strained, dihedrals are observed in ETIM8CHI in the region near position 232, which, is in agreement with the observation that ETIM8CHI and E.coli TIM have, similar stabilities. The largest CA (C-alpha atom) movements, approximately 3 A, are seen for the C-terminal end of helix 8 (associated, with the outward rotation of Phe224) and for the residues in the loop, after helix 1 (associated with sequence changes in helix 8). From the, structure it is not clear why the kcat of ETIM8CHI is 10 times lower than, in wild type E.coli TIM.
About this StructureAbout this Structure
1TMH is a Single protein structure of sequence from Escherichia coli with SO4 as ligand. Active as Triose-phosphate isomerase, with EC number 5.3.1.1 Full crystallographic information is available from OCA.
ReferenceReference
Modular mutagenesis of a TIM-barrel enzyme: the crystal structure of a chimeric E. coli TIM having the eighth beta alpha-unit replaced by the equivalent unit of chicken TIM., Kishan R, Zeelen JP, Noble ME, Borchert TV, Mainfroid V, Goraj K, Martial JA, Wierenga RK, Protein Eng. 1994 Aug;7(8):945-51. PMID:7809033
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