Purification, crystallization and preliminary X-ray diffraction analysis of human enolase-phosphatase E1.
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 A resolution from SeMet-derivative crystals at 100 K using synchrotron radiation. The crystals belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 54.02, b = 57.55, c = 87.32 A. The structure was subsequently solved by the multi-wavelength anomalous diffraction (MAD) phasing method.
Wang
Hui
H
Laboratory of Structural Biology, Tsinghua University, Beijing 100084, People's Republic of China.
Pang
Hai
H
Ding
Yi
Y
Li
Yi
Y
Wu
Xiao'ai
X
Rao
Zihe
Z
D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D009097:Multienzyme Complexes / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013329:Structure-Activity Relationship
C500891:MASA enzyme, human; D009097:Multienzyme Complexes; D011994:Recombinant Proteins