Open AccessStructure of an Escherichia coli N ‐acetyl‐ d ‐neuraminic acid lyase mutant, E192N, in complex with pyruvate at 1.45 Å resolution
Author(s) -
Campeotto Ivan,
Carr Stephen B.,
Trinh Chi H.,
Nelson Adam S.,
Berry Alan,
Phillips Simon E. V.,
Pearson Arwen R.
Publication year - 2009
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309109037403
Subject(s) - neuraminic acid , lyase , escherichia coli , chemistry , mutant , stereochemistry , enzyme , biochemistry , crystallography , sialic acid , gene
The structure of a mutant variant of Escherichia coli N ‐acetyl‐ d ‐neuraminic acid lyase (NAL), E192N, in complex with pyruvate has been determined in a new crystal form. It crystallized in space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 78.3, b = 108.5, c = 148.3 Å. Pyruvate has been trapped in the active site as a Schiff base with the catalytic lysine (Lys165) without the need for reduction. Unlike the previously published crystallization conditions for the wild‐type enzyme, in which a mother‐liquor‐derived sulfate ion is strongly bound in the catalytic pocket, the low‐salt conditions described here will facilitate the determination of further E. coli NAL structures in complex with other active‐site ligands.