Protein-protein interactions (PPIs) are strongly predictive of functional relationships among proteins in virtually all processes that take place in the living cell. Therefore, the comprehensive exploration of interactome networks is one of the major goals in systems biology. The aim of Two Hybrid Technologies: Methods and Protocols is to provide a compendium of state-of-the art protocols for the investigation of binary PPIs with the classical yeast two-hybrid (Y2H) approach, Y2H variants and other in vivo methods for PPI mapping.
Divided into two convenient sections, the first gives a survey of protocols that are currently employed for Y2H high-throughput screens by different expert labs in the field. Rather than detailing the principles of screening, which have been described previously, the focus is on different implementations of Y2H interactome mapping. The second section of the book considers innovative PPI detection methods that have the potential to emerge as alternative high-throughput methodologies.
Written in the highly successful Methods in Molecular Biology series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and accessible, Two Hybrid Technologies: Methods and Protocols supplies researchers with a comprehensive toolbox for the identification of biologically relevant protein interactions.
1. Matrix-Based Yeast Two-Hybrid Screen Strategies and Comparison of Systems Roman Hauser, Thorsten Stellberger, Seesandra V. Rajagopala, and Peter Uetz
2. Array-Based Yeast Two-Hybrid Screens: A Practical Guide Roman Hauser, Thorsten Stellberger, Seesandra V. Rajagopala, and Peter Uetz
3. High Throughput Yeast Two-Hybrid Screening George G. Roberts III, Jodi R. Parrish, Bernardo A. Mangiola, and Russell L. Finley Jr
4. A Stringent Yeast Two-Hybrid Matrix Screening Approach for Protein-Protein Interaction Discovery Josephine Worseck, Arndt Grossmann, Mareike Weimann, Anna Hegele, and Ulrich Stelzl
5. High-Throughput Yeast Two-Hybrid Screening of Complex cDNA Libraries Kerstin Mohr and Manfred Koegl
6. Virus-Human Cell Interactomes Lionel Tafforeau, Chantal Rabourdin-Combe and Vincent Lotteau
7. Interactome Mapping in Malaria Parasites: Challenges and Opportunities Douglas J. LaCount
8. Mapping Interactomes with High Coverage and Efficiency Using the Shifted Transversal Design Xiaofeng Xin, Charles Boone, and Nicolas Thierry-Mieg
9. Assigning Confidence Scores for Protein-Protein Interactions Jingkai Yu, Thilakam Murali, and Russell L. Finley Jr
10. The Intergration and Annotation of the Human Interactome in the UniHI Database Gautam Chaurasia and Matthias Futschik
11. Gene-Centered Yeast One-Hybrid Assays John S. Reece-Hoyes and Albertha J.M. Walhout
12. One- plus Two-hybrid System for the Efficient Selection of Missense Mutant Alleles Defective in Protein-Protein Interactions Ji Young Kim, Ok Gu Park, and Young Chul Lee
13. Investigation of Membrane Protein Interactions Using the Split-Ubiquitin Membrane Yeast Two-Hybrid System Julia Petschnigg, Victoria Wong, Jamie Snider, and Igor Stagljar
14. Application of the Split-Protein Sensor Trp1 to Protein Interaction Discovery in the Yeast Saccharomyces cerevisiae Mandana Rezwan, Nicolas Lentze, Lukas Baumann and Daniel Auerbach
15. Tetracycline Repressor-Based Mammalian Two-Hybrid Systems Kathryn Moncivais and Zhiwen Jonathan Zhang
16. The Fluorescent Two-Hybrid (F2H) Assay for Direct Analysis of Protein-Protein Interactions in Living Cells Kourosh Zolghadr, Ulrich Rothbauer and Heinrich Leonhardt
17. ArrayMAPPIT: a Screening Platform for Human Protein Interactome Analysis Sam Lievens, Nele Vanderroost, Dieter Defever, Jose Van der Heyden and Jan Tavernier
18. MAPPIT as a High-Throughput Screening Assay for Modulators of Protein-Protein Interactions in HIV and HCV Bertrand Van Schoubroeck, Koen Van Acker, Gery Dams, Dirk Jochmans, Reginald Clayton, Jan Martin Berke, Sam Lievens, Jose Van der Heyden and Jan Tavernier
19. Integrated Measurement of Split TEV and cis-Regulatory Assays Using EXT Encoded Reporter Libraries Anna Botvinik and Moritz J. Rossner