Open AccessPurification, crystallization and preliminary X‐ray diffraction analysis of human enolase‐phosphatase E1
Author(s) -
Wang Hui,
Pang Hai,
Ding Yi,
Li Yi,
Wu Xiao'ai,
Rao Zihe
Publication year - 2005
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s174430910501184x
Subject(s) - crystallography , crystallization , yield (engineering) , enolase , diffraction , x ray crystallography , materials science , resolution (logic) , chemistry , optics , biology , physics , immunohistochemistry , organic chemistry , artificial intelligence , computer science , immunology , metallurgy
Enolase‐phosphatase E1 (MASA) is a bifunctional enzyme in the ubiquitous methionine‐salvage pathway and catalyzes the continuous reaction of 2,3‐diketo‐5‐methylthio‐1‐phosphopentane to yield the acireductone metabolite. Recombinant human E1 enzyme has been crystallized using the hanging‐drop vapour‐diffusion method and diffraction‐quality crystals were grown at 291 K using PEG 4000 as precipitant. Diffraction data were collected to 1.7 Å resolution from SeMet‐derivative crystals at 100 K using synchrotron radiation. The crystals belong to space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 54.02, b = 57.55, c = 87.32 Å. The structure was subsequently solved by the multi‐wavelength anomalous diffraction (MAD) phasing method.