Link A J, Eng J, Schieltz D M, Carmack E, Mize G J, Morris D R, Garvik B M, Yates J R
Department of Molecular Biotechnology, University of Washington, Seattle 98195, USA.
Nat Biotechnol. 1999 Jul;17(7):676-82. doi: 10.1038/10890.
We describe a rapid, sensitive process for comprehensively identifying proteins in macromolecular complexes that uses multidimensional liquid chromatography (LC) and tandem mass spectrometry (MS/MS) to separate and fragment peptides. The SEQUEST algorithm, relying upon translated genomic sequences, infers amino acid sequences from the fragment ions. The method was applied to the Saccharomyces cerevisiae ribosome leading to the identification of a novel protein component of the yeast and human 40S subunit. By offering the ability to identify >100 proteins in a single run, this process enables components in even the largest macromolecular complexes to be analyzed comprehensively.
我们描述了一种快速、灵敏的方法，用于全面鉴定大分子复合物中的蛋白质，该方法利用多维液相色谱（LC）和串联质谱（MS/MS）来分离和裂解肽段。SEQUEST算法依靠翻译后的基因组序列，从碎片离子中推断氨基酸序列。该方法应用于酿酒酵母核糖体，从而鉴定出酵母和人类40S亚基中的一种新型蛋白质成分。通过一次运行就能鉴定出100多种蛋白质，该方法能够对即使是最大的大分子复合物中的成分进行全面分析。
