Split leucine-specific domain of leucyl-tRNA synthetase from the hyperthermophilic bacterium Aquifex aeolicus

	Split leucine-specific domain of leucyl-tRNA synthetase from the hyperthermophilic bacterium Aquifex aeolicus
	Ma, JJ; Zhao, MW; Wang, ED
刊名	BIOCHEMISTRY
	2006
卷号	45 期号:49 页码:14809-14816
通讯作者	Wang, ED (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, State Key Lab Mol Biol, Inst Biochem & Cell Biol, Shanghai 200031, Peoples R China.,edwang@sibs.ac.cn
英文摘要	Leucyl-tRNA synthetase (LeuRS) from Aquifex aeolicus is the only known heterodimer synthetase. It is named LeuRS alpha beta, and its alpha and beta subunits contain 634 and 289 residues, respectively. Like Thermus thermophilus LeuRS, LeuR alpha beta has a large extra domain, the leucine-specific domain, inserted into the catalytic domain. The subunit split site is exactly in the middle of the leucine-specific domain and may have a unique function. Here, a series of mutants of LeuRS alpha beta consisting of either mutated alpha subunits and wild-type beta subunits or wild-type alpha subunits and mutated beta subunits were constructed and purified. ATP-PPi exchange and aminoacylation activities and the ability of the mutants to charge minihelix(Leu) were assayed. Interaction of the mutants with the tRNA was assessed by gel shift. Two peptides of eight and nine amino acid residues in the domain located in the alpha subunit were found to be essential for the enzyme's activity. We also showed that the domain in LeuRS alpha beta plays an important role in minihelixLeu recognition. Additionally, the domain was found to have little impact on the assembly of the heterodimer, to play a role in the thermal stability of the whole enzyme, and to interact with the cognate tRNA in the predicted manner.
学科主题	Biochemistry & Molecular Biology
类目[WOS]	Biochemistry & Molecular Biology
关键词[WOS]	ESCHERICHIA-COLI ; CRYSTAL-STRUCTURE ; BETA-SUBUNIT ; PURIFICATION ; TRNA(LEU) ; DISCRIMINATION ; EXPRESSION ; COMPLEX
收录类别	SCI
语种	英语
WOS记录号	WOS:000242516100026
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/1809]
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	Ma, JJ,Zhao, MW,Wang, ED. Split leucine-specific domain of leucyl-tRNA synthetase from the hyperthermophilic bacterium Aquifex aeolicus[J]. BIOCHEMISTRY,2006,45(49):14809-14816.
APA	Ma, JJ,Zhao, MW,&Wang, ED.(2006).Split leucine-specific domain of leucyl-tRNA synthetase from the hyperthermophilic bacterium Aquifex aeolicus.BIOCHEMISTRY,45(49),14809-14816.
MLA	Ma, JJ,et al."Split leucine-specific domain of leucyl-tRNA synthetase from the hyperthermophilic bacterium Aquifex aeolicus".BIOCHEMISTRY 45.49(2006):14809-14816.