Crystallization and preliminary crystallographic analysis of nosiheptide-resistance methyltransferase from Streptomyces actuosus in complex with SAM.
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 .
Yang
Huirong
H
Department of Pathology, Cancer Hospital, Fudan University, Shanghai 200032, People's Republic of China.
Wang
Ping
P
Dong
Zhenghong
Z
Li
Xueyuan
X
Gong
Rui
R
Yang
Ying
Y
Li
Ze
Z
Xu
Youwei
Y
Xu
Yanhui
Y
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
D013844:Thiazoles; D012436:S-Adenosylmethionine; D008780:Methyltransferases; C014134:nosiheptide
Antimicrob Agents Chemother. 2000 Feb;44(2):411-3
10639372
J Antibiot (Tokyo). 1978 Jun;31(6):623-5
681244
Mol Cell. 2008 Apr 11;30(1):26-38
18406324
Eur J Biochem. 1994 Sep 1;224(2):431-7
7925357
J Biol Chem. 2009 Jun 19;284(25):17013-17020
19369248
Methods Enzymol. 1997;276:307-26
27754618
J Bacteriol. 1980 May;142(2):455-61
6155371
Microbiol Rev. 1994 Mar;58(1):10-26
8177168
J Mol Biol. 2005 Jan 21;345(3):535-45
15581897
Nucleic Acids Res. 2008 Jan;36(Database issue):D281-8
18039703
Cell. 1999 May 14;97(4):491-502
10338213
Gene. 1990 Jul 2;91(1):9-17
2401410
Science. 1991 Oct 4;254(5028):51-8
1925561
Phytochemistry. 2006 Aug;67(15):1686-98
16766004
Structure. 2002 Oct;10(10):1303-15
12377117
Chem Biol. 2003 Aug;10(8):769-78
12954336
Microbiol Rev. 1994 Jun;58(2):211-32
7521510
Nature. 1977 Jan 13;265(5590):189-90
834263
Gene. 1993 May 15;127(1):63-9
8486289
J Anim Sci. 1984 Nov;59(5):1125-8
6511687