Abstract
| I. | Introduction | 00 |
| II. | The Yeast Two‐Hybrid System | 00 |
| | A. A Brief History | 00 |
| | 1. Basis of Yeast Two‐Hybrid System | 00 |
| | 2. Development of the System | 00 |
| | B. Current Systems | 00 |
| | 1. The GAL4 and LexA Systems | 00 |
| | 2. The Sos Recruitment System | 00 |
| | 3. The Split‐Ubiquitin System | 00 |
| | 4. The Dual‐Bait System | 00 |
| | 5. Other Two‐Hybrid Systems | 00 |
| | 6. Variations on a Theme | 00 |
| | C. Applications of Yeast Two‐Hybrid | 00 |
| III. | Studying Protein Interactions by MS | 00 |
| IV. | Genome‐Wide Analysis of Protein–Protein Interactions | 00 |
| | A. Yeast Two‐Hybrid | 00 |
| | B. Mass Spectrometry | 00 |
| | C. Comparison of the Two Approaches | 00 |
| | D. Placing Interactions in Context | 00 |
| V. | Emerging Technologies | 00 |
| VI. | Conclusions | 00 |
| Acknowledgments | 00 |
| Abbreviations | 00 |
| References | 00 |
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