流感嗜血杆菌tRNA（m1G37）甲基转移酶的结晶及初步X射线晶体学分析

Crystallization and preliminary X-ray crystallographic analysis of tRNA(m1G37)methyltransferase from Haemophilus influenzae.

作者信息

Kim Hyeon-Woo, Ahn Hyung Jun, Yoon Hye-Jin, Kim Hyung-Wook, Baek Seung-Hun, Suh Se Won

机构信息

Structural Proteomics Laboratory, School of Chemistry and Molecular Engineering, Seoul National University, Seoul 151-742, South Korea.

出版信息

Acta Crystallogr D Biol Crystallogr. 2003 Jan;59(Pt 1):183-4. doi: 10.1107/s0907444902019716. Epub 2002 Dec 19.

DOI:10.1107/s0907444902019716

PMID:12499565

Abstract

The enzyme tRNA(m(1)G37)methyltransferase (TrmD) catalyzes the transfer of a methyl group from S-adenosyl-L-methionine (AdoMet) specifically to guanosine at position 37 within a subset of tRNA species in bacteria. The modified guanosine is next to the anticodon and is important for the maintenance of the correct reading frame during translation. TrmD from Haemophilus influenzae with both N- and C-terminal tags was overexpressed in Escherichia coli and crystallized at 297 K using sodium acetate as a precipitant. Native X-ray diffraction data were collected to 1.85 A resolution. The crystals are rhombohedral, belonging to the space group R32, with unit-cell parameters a = b = 98.05, c = 176.79 A, alpha = beta = 90, gamma = 120 degrees. The presence of one monomer of recombinant TrmD in the crystallographic asymmetric unit gives a V(M) of 3.07 A(3) Da(-1) and a solvent content of 59.9%.

摘要

tRNA（m(1)G37）甲基转移酶（TrmD）催化将S-腺苷-L-甲硫氨酸（AdoMet）上的甲基特异性转移至细菌中特定tRNA种类的37位鸟苷上。修饰后的鸟苷紧邻反密码子，对翻译过程中维持正确的阅读框很重要。带有N端和C端标签的流感嗜血杆菌TrmD在大肠杆菌中过表达，并以乙酸钠作为沉淀剂于297 K下结晶。收集到了分辨率为1.85 Å的天然X射线衍射数据。晶体为菱面体，属于R32空间群，晶胞参数a = b = 98.05，c = 176.79 Å，α = β = 90°，γ = 120°。晶体学不对称单元中存在一个重组TrmD单体，其V(M)为3.07 Å³ Da⁻¹，溶剂含量为59.9%。