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Purification, crystallization and preliminary X‐ray diffraction studies of UDP‐glucose:tetrahydrobiopterin α‐glucosyltransferase (BGluT) from Synechococcus sp. PCC 7942
Author(s) -
Killivalavan Asaithambi,
Zhuang Ningning,
Park Young Shik,
Lee Kon Ho
Publication year - 2014
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x13034298
Subject(s) - chemistry , glucosyltransferase , tetrahydrobiopterin , polyethylene glycol , crystallization , crystallography , moiety , synechococcus , chromatography , size exclusion chromatography , enzyme , biochemistry , stereochemistry , cyanobacteria , cofactor , biology , organic chemistry , bacteria , genetics
A UDP‐glucose:tetrahydrobiopterin α‐glucosyltransferase (BGluT) enzyme was discovered in the cyanobacterium Synechococcus sp. PCC 7942 which transfers a glucose moiety from UDP‐glucose to tetrahydrobiopterin (BH 4 ). BGluT protein was overexpressed with selenomethionine labelling for structure determination by the multi‐wavelength anomalous dispersion method. The BGluT protein was purified by nickel‐affinity and size‐exclusion chromatography. It was then crystallized by the hanging‐drop vapour‐diffusion method using a well solution consisting of 0.1 M bis‐tris pH 5.5, 19%( w / v ) polyethylene glycol 3350 with 4%( w / v ) D(+)‐galactose as an additive. X‐ray diffraction data were collected to 1.99 Å resolution using a synchrotron‐radiation source. The crystals belonged to the monoclinic space group C 2, with unit‐cell parameters a = 171.35, b = 77.99, c = 53.77 Å, β = 90.27°.