title: Crystallization and preliminary crystallographic analysis of nosiheptide-resistance methyltransferase from Streptomyces actuosus in complex with SAM.
abstract: Nosiheptide-resistance methyltransferase (NSR) methylates 23S rRNA at the nucleotide adenosine 1067 in Escherichia coli and thus contributes to resistance against nosiheptide, a sulfur-containing peptide antibiotic. Here, the expression, purification and crystallization of NSR from Streptomyces actuosus are reported. Diffracting crystals were grown by the hanging-drop vapour-diffusion method in reservoir solution consisting of 0.35 M ammonium chloride, 24%(w/v) PEG 3350, 0.1 M MES pH 5.7 at 293 K. Native data have been collected from the apo enzyme and a SAM complex, as well as apo SeMet SAD data. The diffraction patterns of the apo form of NSR, of NSR complexed with SAM and of SeMet-labelled NSR crystals extended to 1.90, 1.95 and 2.25 A resolution, respectively, using synchrotron radiation. All crystals belonged to space group P2(1), with approximate unit-cell parameters a = 64.6, b = 69.6, c = 64.9 A, beta = 117.8 degrees .
authors.affiliation: Department of Pathology, Cancer Hospital, Fudan University, Shanghai 200032, People's Republic of China.
authors.forename: Huirong
authors.initials: H
authors.lastname: Yang
authors.forename: Ping
authors.initials: P
authors.lastname: Wang
authors.forename: Zhenghong
authors.initials: Z
authors.lastname: Dong
authors.forename: Xueyuan
authors.initials: X
authors.lastname: Li
authors.forename: Rui
authors.initials: R
authors.lastname: Gong
authors.forename: Ying
authors.initials: Y
authors.lastname: Yang
authors.forename: Ze
authors.initials: Z
authors.lastname: Li
authors.forename: Youwei
authors.initials: Y
authors.lastname: Xu
authors.forename: Yanhui
authors.initials: Y
authors.lastname: Xu
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray; D024881:Drug Resistance, Bacterial; D008780:Methyltransferases / Q000737:chemistry* / Q000378:metabolism; D012436:S-Adenosylmethionine / Q000737:chemistry* / Q000378:metabolism; D013302:Streptomyces / Q000187:drug effects / Q000201:enzymology*; D013844:Thiazoles / Q000494:pharmacology
chemical_list: D013844:Thiazoles; D012436:S-Adenosylmethionine; D008780:Methyltransferases; C014134:nosiheptide
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