Mass spectra of partial protein hydrolysates as a multiple phase check for long polypeptides deduced from DNA sequences: NH2-terminal segment of alanine tRNA synthetase.
Mass spectra of partial protein hydrolysates as a multiple phase check for long polypeptides deduced from DNA sequences: NH2-terminal segment of alanine tRNA synthetase.
复制标题
部分蛋白质水解物的质谱作为对从 DNA 序列推导出的长多肽的多相检查：丙氨酸 tRNA 合成酶的 NH2 末端片段。
DOI：
10.1073/pnas.77.11.6531
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发表时间：
1980
影响因子：
11.1
通讯作者：
Schimmel,PR
中科院分区：
文献类型：
--
作者：
Herlihy,WC;Royal,NJ;Biemann,K;Putney,SD;Schimmel,PR
已经开发了一种用于快速和准确测定大蛋白质的氨基酸序列的策略，例如被称为氨酰tRNA合成酶的蛋白质类的许多成员。该策略包括将感兴趣的蛋白质的基因的DNA测序与整个蛋白质或其非常大的片段的部分水解产物中的四肽和五肽的气相色谱质谱鉴定相结合。这些肽与散布在整个结构基因中的密码子块相匹配。四肽和五肽序列足够长，它们不太可能在蛋白质序列中重复或出现在不正确的阅读框架中;因此，它们可以被放置在DNA上独特的密码子簇中。该方法严格地建立了贯穿结构基因的整个长度的DNA的适当定相，并且由此从DNA序列准确地读取蛋白质序列。这种方法被用于确定大肠杆菌丙氨酸tRNA合成酶的氨基酸序列，这种蛋白质具有大约900个氨基酸。本文报道了NH_2末端前165个氨基酸的序列。
A strategy has been developed for rapid and accurate determination of the amino acid sequence of large proteins, such as many of the members of the class of proteins known as aminoacyl tRNA synthetases. This strategy involves combining DNA sequencing of the gene for the protein of interest with gas chromatographic mass spectrometric identification of tetra- and pentapeptides in partial hydrolysates of the entire protein or very large fragments thereof. These peptides are matched to blocks of codons at locations scattered throughout the entire structural gene. Tetra- and pentapeptide sequences are sufficiently long that they are unlikely to be repeated in the protein sequence or to occur in an incorrect reading frame; therefore, they can be placed at unique clusters of codons on the DNA. This procedure rigorously establishes the proper phasing of the DNA throughout the entire length of the structural gene, and the protein sequence is thereby accurately read from the DNA sequence. This approach is being used to determine the amino acid sequence of EScherichia coli alanine tRNA synthetase, a protein that has approximately 900 amino acids. This paper reports the sequence of the first 165 amino acids from the NH2 terminus.