Direct analysis of protein complexes using mass spectrometry.

Link AJ, Eng J, Schieltz DM, Carmack E, Mize GJ, Morris DR, Garvik BM, Yates JR
Nat Biotechnol. 1999 17 (7): 676-82

PMID: 10404161 · 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.

MeSH Terms (10)

Algorithms Amino Acid Sequence Chromatography, Liquid Humans Mass Spectrometry Molecular Sequence Data Ribosomal Proteins Ribosomes Saccharomyces cerevisiae Sensitivity and Specificity

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