Probing the potential metal binding site in Escherichia coli 3‐deoxy‐d‐arabino‐heptulosonate 7‐phosphate synthase (phenylalanine‐sensitive)

FEBS Letters

Sundaram, Appavu K¹  Howe, David L¹  Sheflyan, Galina Ya¹  Woodard, Ronald W¹ 

[1] Interdepartmental Program in Medicinal Chemistry, College of Pharmacy, University of Michigan, Ann Arbor, MI 48109-1065, USA

关键词: Deoxy-d-arabino-heptulosonate 7-phosphate synthase;    Metal binding;    Phosphoenolpyruvate;    Erythrose 4-phosphate;    Site-directed mutagenesis;    BTP;    1;    3-bis[tris-(hydroxymethyl)-methylamino]propane;    DAH 7-P;    3-deoxy-d-arabino-heptulosonic acid 7-phosphate;    DAH 7-P synthase;    3-deoxy-d-arabino-heptulosonic acid 7-phosphate synthase;    E 4-P;    erythrose 4-phosphate;    EDTA;    [ethylenebis(oxyethylenenitrilo)]tetraacetic acid;    PEP;    phosphoenolpyruvate;   

DOI  :  10.1016/S0014-5793(98)01545-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The active site residues of the proposed metal binding site of DAH 7-P synthase (phe) were probed by site-directed mutagenesis of C61 to glycine and serine, H64 to glycine, and with the double mutant C61H/H64C. While C61S and C61H/H64C were inactive, both C61G and H64G were active. All mutants, regardless of enzymatic activity, bound one equivalent of Fe²⁺ per monomeric unit. Even though C61 and H64 were shown not to be metal ligands for the DAH 7-P synthase (phe), they may provide some of the backbone interactions/secondary structural elements necessary to properly form the metal binding pocket.

【 授权许可】

Unknown