Mass spectrometry-based shotgun proteomic analysis of C. elegans protein complexes

Bryan R Fonslow, James J Moresco, Patricia G Tu, Antti P Aalto, Amy E Pasquinelli, Andrew G Dillin, John R Yates

Research output: Contribution to journalReview ArticleScientificpeer-review

8 Citations (Scopus)

Abstract

Mass spectrometry (MS)-based shotgun proteomics is an enabling technology for the study of C. elegans proteins. When coupled with co-immunoprecipitation (CoIP), new interactions and functions among proteins can be discovered. We provide a general background on protein complexes and methods for their analysis, along with the lifecycle and interaction types of proteins that ultimately define the identifiable components of protein complexes. We highlight traditional biochemical methods to evaluate whether the complexes are sufficiently pure and abundant for analysis with shotgun proteomics. We present two CoIP-MS case studies of protein complexes from C. elegans, using both endogenous and fusion protein antibodies to illustrate the important aspects of their analyses. We discuss results from mass spectrometers with differences in mass accuracy and resolution, along with the relevant information that can be extracted from the data generated, such as protein relative abundance, post-translational modifications, and identification confidence. Finally, we illustrate how comparative analysis can reveal candidate binding partners for biological follow-up and validation. This chapter should act as a complement and extension to the WormBook chapter Biochemistry and molecular biology, which describes tandem affinity purification (TAP) of protein complexes for analysis by mass spectrometry.

Original languageEnglish
Pages (from-to)1-18
Number of pages18
JournalWormBook: the online review of C. elegans biology
DOIs
Publication statusPublished - 24 Jun 2014
MoE publication typeA2 Review article in a scientific journal

Keywords

  • Animals
  • Caenorhabditis elegans/genetics
  • Caenorhabditis elegans Proteins/genetics
  • Immunoprecipitation
  • Mass Spectrometry
  • Multiprotein Complexes
  • Protein Processing, Post-Translational
  • Proteome
  • Proteomics/methods

Fingerprint

Dive into the research topics of 'Mass spectrometry-based shotgun proteomic analysis of C. elegans protein complexes'. Together they form a unique fingerprint.

Cite this