Methionyl-tRNA synthetase from Escherichia coli. Primary structure of the active crystallised tryptic fragment.

Methionyl-tRNA synthetase from Escherichia coli. Primary structure of the active crystallised tryptic fragment.
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来自大肠杆菌的甲硫氨酰-tRNA 合成酶。

DOI:
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发表时间:
2005
期刊:
European Journal of Biochemistry
影响因子:
--
通讯作者:
C. Bruton
C. Bruton
中科院分区:
--
文献类型:
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作者:
D. Barker;J. Ebel;R. Jakes;C. Bruton

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克隆到 pBR322 中的大肠杆菌染色体 DNA 的 3300 个碱基片段将补充甲硫氨酸营养缺陷型,其中病变是有缺陷的甲硫氨酰-tRNA 合成酶,对甲硫氨酸的亲和力大大降低。这些转化体的粗提物含有升高水平的蛋白质,该蛋白质的亚基分子量为66 000,具有体外甲硫氨酰-tRNA合成酶氨酰化活性,并且与抗(甲硫氨酰-tRNA合成酶)抗体发生交叉反应。该多肽比已充分表征的甲硫氨酰-tRNA 合成酶的胰蛋白酶片段稍大一些。克隆插入片段一端的 1750 个残基的 DNA 序列编码一个非终止的开放阅读框,我们可以在其中找到大量的甲硫氨酰-tRNA 合成酶胰蛋白酶肽和胰凝乳蛋白酶肽。我们还对这个编码片段上游的 300 个核苷酸进行了测序,我们在推定的甲硫氨酰-tRNA 合成酶启动子区域发现了一个大的反向重复序列。
A 3300-base segment of Escherichia coli chromosomal DNA, cloned into pBR322, will complement a methionine auxotroph in which the lesion is a defective methionyl-tRNA synthetase with a much reduced affinity for methionine. Crude extracts of these transformants contain elevated levels of a protein which has a subunit molecular weight of 66 000, methionyl-tRNA synthetase aminoacylation activity in vitro and which cross-reacts with anti-(methionyl-tRNA synthetase) antibodies. This polypeptide is very slightly larger than the well-characterised and crystallised tryptic fragment of methionyl-tRNA synthetase. A DNA sequence of 1750 residues at one end of the cloned insert codes for a non-terminated open reading frame in which we can locate a large number of methionyl-tRNA synthetase tryptic and chymotryptic peptides. We have also sequenced 300 nucleotides upstream of this coding segment where we find a large invert repeat in the putative methionyl-tRNA synthetase promoter region.
大型氨酰基-tRNA 合成酶的一级结构。
DOI: 10.1126/science.7025207
发表时间: 1981
期刊: Science (New York, N.Y.)
影响因子: --
作者:
Putney,SD;Royal,NJ;NeumandeVegvar,H;Herlihy,WC;Biemann,K;Schimmel,P
通讯作者: Schimmel,P