title: Purification, crystallization and preliminary X-ray diffraction studies of UDP-glucose:tetrahydrobiopterin α-glucosyltransferase (BGluT) from Synechococcus sp. PCC 7942.
abstract: 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 (BH4). 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 C2, with unit-cell parameters a = 171.35, b = 77.99, c = 53.77 Å, β = 90.27°.
authors.affiliation: Department of Microbiology, School of Medicine, Gyeongsang National University, Jinju 660-751, Republic of Korea.
authors.forename: Asaithambi
authors.initials: A
authors.lastname: Killivalavan
authors.affiliation: Plant Molecular Biology and Biotechnology Research Center (PMBBRC), Gyeongsang National University, Jinju 660-701, Republic of Korea.
authors.forename: Ningning
authors.initials: N
authors.lastname: Zhuang
authors.affiliation: School of Biological Sciences, Inje University, Kimhae 621-749, Republic of Korea.
authors.forename: Young Shik
authors.initials: YS
authors.lastname: Park
authors.affiliation: Department of Microbiology, School of Medicine, Gyeongsang National University, Jinju 660-751, Republic of Korea.
authors.forename: Kon Ho
authors.initials: KH
authors.lastname: Lee
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification; D001483:Base Sequence; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D017931:DNA Primers; D005964:Glucosyltransferases / Q000737:chemistry* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D011487:Protein Conformation; D046940:Synechococcus / Q000201:enzymology*
chemical_list: D001426:Bacterial Proteins; D017931:DNA Primers; C437230:BGluT protein, Synechococcus sp. PCC 7942; D005964:Glucosyltransferases
keywords: glucosyltransferase; pteridine glycosyltransferase; tetrahydrobiopterin