The effect of N-terminal changes on arginyl-tRNA synthetase from Escherichia coli

	The effect of N-terminal changes on arginyl-tRNA synthetase from Escherichia coli
	Liu, W; Liu, MF; Xia, M; Wang, YL
刊名	ACTA BIOCHIMICA ET BIOPHYSICA SINICA
	2002
卷号	34 期号:2 页码:131-137
关键词	arginyl-tRNA synthetase N-terminal activity mutatation
通讯作者	,
英文摘要	An Asn(2) deleted mutant of Escherichia coli arginyl-tRNA synthetase deleted Asn2 and a chimera mutant, in which the N-terminal 23 amino acid residues of yeast arginyl-tRNA synthetase were appended to the N-terminus of Escherichia coli synthetase, were synthesized and studied. The expression of the deletion and chimera mutants in Escherichia coli formed inclusion bodies, presumably due to improper folding of the proteins. Relative to the native enzyme, the deletion mutant showed full amino acid activation activity and a 26% reduction in aminoacylation activity, while the chimera mutant lost 93% and 96% activities in amino acid activation and aminoacylation, respectively, and did not aminoacylate yeast tRNA(Arg) at all. The mutant deleted Asn(2) and Ile(3) was able to be expressed in Escherichia coli but not stable to be purified. The emission maximum wavelength in the fluorescence spectra of the chimera mutants shifted to longer one and the corresponding intensities decreased, when compared with those of the native enzyme. The data show that the conformation of the mutants are different and the tryptophan residues in the mutants are more exposed than those in the native enzyme. An estimate of the secondary structure of the mutant enzymes from their far ultraviolet CID spectra showed that the chimera mutant contained less alpha-helix, more beta-sheet and slightly higher fraction of random coil, as compared with the native enzyme. The results indicate that an intact N-terminal domain of E. coli arginyl-tRNA synthetase is important to its activity and correct folding.
学科主题	Biochemistry & Molecular Biology; Biophysics
类目[WOS]	Biochemistry & Molecular Biology ; Biophysics
关键词[WOS]	TRANSFER-RNA-SYNTHETASE ; CIRCULAR-DICHROISM ; AMINOACYLATION ; PURIFICATION ; COMPLEX ; DOMAIN ; OVERPRODUCTION ; RECOGNITION ; RESIDUES ; DELETION
收录类别	SCI
语种	英语
WOS记录号	WOS:000174549400003
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/2446]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Liu, W,Liu, MF,Xia, M,et al. The effect of N-terminal changes on arginyl-tRNA synthetase from Escherichia coli[J]. ACTA BIOCHIMICA ET BIOPHYSICA SINICA,2002,34(2):131-137.
APA	Liu, W,Liu, MF,Xia, M,&Wang, YL.(2002).The effect of N-terminal changes on arginyl-tRNA synthetase from Escherichia coli.ACTA BIOCHIMICA ET BIOPHYSICA SINICA,34(2),131-137.
MLA	Liu, W,et al."The effect of N-terminal changes on arginyl-tRNA synthetase from Escherichia coli".ACTA BIOCHIMICA ET BIOPHYSICA SINICA 34.2(2002):131-137.