Two-hybrid systems : methods and protocols

書誌事項

Two-hybrid systems : methods and protocols

edited by Paul N. MacDonald

(Methods in molecular biology / John M. Walker, series editor, v. 177)

Humana Press, c2001

  • : hardcover
  • : comb

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注記

Includes bibliographical references and index

内容説明・目次

内容説明

Many, if not all, essential biological processes require selective interactions between proteins. Complex signaling systems require sequential, ordered protein-protein interactions at essentially all levels of the signaling cascade. For example, peptide hormones interact with selective membrane receptor proteins, and autophosphorylation of the receptor then recruits other key regulatory proteins that initiate kinase cascades in which each phosphorylation event requires selective recognition of the protein substrate. The ultimate signaling effect, in many cases, is the regulation of RNA polymerase II-directed transcr- tion in the nucleus, a process that involves numerous, multiprotein complexes important for transcription initiation, elongation, termination, and reinitiation. Defining, characterizing, and understanding the relevance of these protein- protein interactions is an arduous task, but substantial inroads have been made over the past 20 years. The development of more recent methodologies, such as mammalian expression systems, immunopurification schemes, expression cloning strategies, surface plasmon resonance (BiaCore), and nanosequencing technologies, has contributed a wealth of new insights into these complex multiprotein mechanisms and clearly accelerated the discovery process. Arguably, the yeast two-hybrid system has been one of the predominant and most powerful tools in this discovery process.

目次

Part I. General Methods The Two-Hybrid System: A Personal View Stanley Fields and Paul L. Bartel Growth and Maintenance of Yeast Lawrence W. Bergman Media Formulations for Various Two-Hybrid Systems Michael Saghbini, Denise Hoekstra, and Jim Gautsch Yeast Two-Hybrid Vectors and Strains Philip James High-Efficiency Transformation of Plasmid DNA into Yeast Robin A. Woods and R. Daniel Gietz Qualitative and Quantitative Assessment of Interactions Monica M. Montano Strategies for Rescuing Plasmid DNA from Yeast Two-Hybrid Colonies, Alyson Byrd and Rene St-Arnaud Part II. False Positives Two-Hybrid System and False Positives: Approaches to Detection and Elimination Ilya G. Serebriiskii and Erica A. Golemis Confirming Yeast Two-Hybrid Protein Interactions Using In Vitro Glutathione-S-Transferase Pulldowns Dennis M. Kraichely and Paul N. MacDonald Two-Hybrid Interactions Confirmed by Coimmunoprecipitation of Epitope-Tagged Clones Louie Naumovski Part III. Applications Smad Interactors in Bone Morphogenetic Protein Signaling Xiangli Yang and Xu Cao Protein Interactions Important in Eukaryotic Translation Initiation Katsura Asano and Alan G. Hinnebusch Steroid Receptor and Ligand-Dependent Interaction with Coactivator Proteins Sergio A. Onate Interaction of Cellular Apoptosis Regulating Proteins with Adenovirus Anti-Apoptosis Protein E1B-19K Thirugnana Subramanian and G. Chinnadurai Part IV. Alternative Strategies Mammalian Two-Hybrid Assays: Analyzing Protein-Protein Interactions in the Transforming Growth Factor-_ Signaling Pathway Xin-Hua Feng and Rik Derynck One-Hybrid Systems for Detecting Protein-DNA Interactions Mary Kate Alexander, Brenda D. Bourns, and Virginia A.Zakian The Split-Hybrid System: Uncoding Multiprotein Networks and Defining Mutations That Affect Protein Interactions Phyllis S. Goldman, Anthony J. DeMaggio, Richard H. Goodman, and Merl F. Hoekstra Three-Hybrid Screens: Inducible Third-Party Systems Bjoern Sandrock, Franck Tirode, and Jean-Marc Egly Three-Hybrid Screens for RNA-Binding Proteins: Proteins Binding 3' End of Histone mRNA Zbigniew Dominski and William F. Marzluff Membrane Recruitment Systems for Analysis of Protein-Protein Interactions Ami Aronheim

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