Studying the interactome with the yeast two-hybrid system and mass spectrometry

Authors


Abstract

  I.Introduction00
 II.The Yeast Two-Hybrid System00
 A.  A Brief History00
     1.  Basis of Yeast Two-Hybrid System00
     2.  Development of the System00
 B.  Current Systems00
     1.  The GAL4 and LexA Systems00
     2.  The Sos Recruitment System00
     3.  The Split-Ubiquitin System00
     4.  The Dual-Bait System00
     5.  Other Two-Hybrid Systems00
     6.  Variations on a Theme00
 C.  Applications of Yeast Two-Hybrid00
  III.Studying Protein Interactions by MS00
 IV.Genome-Wide Analysis of Protein–Protein Interactions00
 A.  Yeast Two-Hybrid00
 B.  Mass Spectrometry00
 C.  Comparison of the Two Approaches00
 D.  Placing Interactions in Context00
  V.Emerging Technologies00
 VI.Conclusions00
Acknowledgments00
Abbreviations00
References00

Protein interactions are crucial to the life of a cell. The analysis of such interactions is allowing biologists to determine the function of uncharacterized proteins and the genes that encode them. The yeast two-hybrid system has become one of the most popular and powerful tools to study protein–protein interactions. With the advent of proteomics, the two-hybrid system has found a niche in interactome mapping. However, it is clear that only by combining two-hybrid data with that from complementary approaches such as mass spectrometry (MS) can the interactome be analyzed in full. This review introduces the yeast two-hybrid system to those unfamiliar with the technique, and discusses how it can be used in combination with MS to unravel the network of protein interactions that occur in a cell. © 2004 Wiley Periodicals, Inc., Mass Spec Rev

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