The Cross-and-Capture system: a versatile tool in yeast proteomics

Methods. 2012 Dec;58(4):360-6. doi: 10.1016/j.ymeth.2012.07.017. Epub 2012 Jul 24.

Abstract

High-throughput technologies such as affinity purification and yeast two-hybrid (Y2H) screening have been applied to explore protein-protein interactions (PPIs) in the model organism Saccharomyces cerevisiae. The "Cross-and-Capture" system is an alternative approach for an assessment of PPIs using two arrayed collections of differentially tagged yeast strains. In its current implementation, Cross-and-Capture encompasses ∼500 endogenously tagged yeast proteins, predominantly with roles in DNA metabolism and maintenance. The tagged arrays can also serve other purposes in yeast proteomics, such as monitoring of protein expression and the detection of posttranslational protein modifications. In this article, I summarize the development of this tool and describe its application to detect protein complexes and to screen for novel PPIs.

MeSH terms

  • Base Sequence
  • Blotting, Western
  • Chromatography, Affinity
  • Crosses, Genetic
  • Genetic Engineering
  • Open Reading Frames
  • Protein Binding
  • Protein Interaction Mapping / methods*
  • Proteomics
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / isolation & purification
  • Saccharomyces cerevisiae Proteins / metabolism*

Substances

  • Saccharomyces cerevisiae Proteins