Crystallization and 1.1 Angstrom Diffraction of Chorismate Lyase From Escherichia Coli

Published

February 1, 2000

Author(s)

C Stover, M P. Mayhew, Marcia J. Holden, A Howard, David Travis Gallagher

Abstract

Clorismate pathway enzymes are important as producers of nonnucleotide aromatic compounds. The enzyme chorismate layase from Escherichia coli has been crystallized in four distinct forms, three of which have been characterized by x-ray diffraction. Despite widespread screening, all four crystal forms grow from the same chemical conditions. The wild-type enzyme tends to aggregate, even in the presence of a reducing agent, and yielded only one crystal form (monoclinic, form 1) that grew in intricate clusters. Chemical modification of the cysteines mitgated problems with aggregation and solubility but did not affect crystal growth behavior. Protein aggregation was largely eliminated by mutating the protein s two cysteines to serines. The double mutant retains full enzymatic activity and crystallizes in three new forms, one of which (triclinic) diffracts to 1.1 resolution.

Citation

Journal of Structural Biology

Volume

129

Issue

1

Pub Type

Journals

Keywords

chemical modification chorismic, chorismic acid, crystal forms, protein crystal growth, site-directed mutagenesis

Citation

Stover, C. , Mayhew, M. , Holden, M. , Howard, A. and Gallagher, D. (2000), Crystallization and 1.1 Angstrom Diffraction of Chorismate Lyase From Escherichia Coli, Journal of Structural Biology (Accessed July 16, 2026)

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