从聚丙烯酰胺凝胶上洗脱下来的蛋白质的内部序列分析。

Internal sequence analysis of proteins eluted from polyacrylamide gels.

作者信息

Hwang B J, Smith A J, Chu G

机构信息

Department of Medicine, Stanford University Medical Center, CA 94305, USA.

出版信息

J Chromatogr B Biomed Appl. 1996 Nov 15;686(2):165-75. doi: 10.1016/s0378-4347(96)00230-7.

DOI:10.1016/s0378-4347(96)00230-7

PMID:8971597

Abstract

We have developed an elution-digestion-sequencing (EDS) method, which yields the internal amino acid sequence of partially purified proteins. The overall yield for the method was greater than 60%. The method yielded peptide peaks that could be sequenced on HPLC for all tested proteins with masses from 45 to 200.10(3) and yielded internal amino acid sequence information when as little as 10 pmol of partially purified protein was used as the starting material. The EDS method was extremely reliable and gave sequence information for each of 25 proteins tested, including high-molecular-mass proteins (M(r) > 100.10(3)) that were difficult to sequence by other methods.

摘要

我们开发了一种洗脱-消化-测序（EDS）方法，该方法可产生部分纯化蛋白质的内部氨基酸序列。该方法的总产率大于60%。对于所有质量在45至200.10(3)之间的测试蛋白质，该方法产生的肽峰可在高效液相色谱（HPLC）上进行测序，并且当仅使用10皮摩尔的部分纯化蛋白质作为起始材料时，就能获得内部氨基酸序列信息。EDS方法极其可靠，为所测试的25种蛋白质中的每一种都提供了序列信息，包括其他方法难以测序的高分子量蛋白质（M(r)>100.10(3)）。