Mass spectrometry and EST-database searching allows characterization of the multi-protein spliceosome complex.

G Neubauer, A King, J Rappsilber, C Calvio, M Watson, P Ajuh, Judith Elizabeth Sleeman, A Lamond, M Mann

Research output: Contribution to journalArticlepeer-review

407 Citations (Scopus)

Abstract

Many important cell mechanisms are carried our and regulated by multi-protein complexes, for example, transcription and RNA processing machinery, receptor complexes end cytoskeletal structures. Most of these complexes remain only partially characterized due to the difficulty of conventional protein analysis methods. The rapid expansion of DNA sequence databases now provides whole or partial gene sequences of model organisms, and recent advances in protein microcharacterization via mass spectrometry allow the possibility of linking these DNA sequences to the proteins in functional complexes(1). This approach has been demonstrated in organisms whose genomes have been sequenced(2), such as budding yeast. Here we report the first characterization of an entire mammalian multi-protein complex using these methods. The machinery that removes introns from mRNA percursors-the spliceosome-is a large multi-protein complex(3,4). Approximately half of the components excised from a two-dimensional gel separation of the spliceosome were found in protein sequence databases. Using nanoelectrospray mass spectrometry, the remainder were identified and cloned using public expressed sequence tag (EST) databases. Existing EST databases are thus already sufficiently complete to allow rapid characterization of large mammalian protein complexes via mass spectrometry.

Original languageEnglish
Pages (from-to)46-50
Number of pages5
JournalNature Genetics
Volume20
Issue number1
DOIs
Publication statusPublished - Sept 1998

Keywords

  • POLYACRYLAMIDE GELS
  • IDENTIFICATION
  • GENOME
  • NANOELECTROSPRAY
  • HELICASE
  • U4/U6
  • BOX
  • U1

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