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Two-Hybrid Systems: Methods and Protocols

Two-Hybrid Systems: Methods and Protocols

Paperback

Series: Methods in Molecular Biology, Book 177

Medical ReferenceBiologyChemistry

ISBN10: 0896038084
ISBN13: 9780896038080
Publisher: Springer Nature
Published: May 17 2001
Pages: 336
Weight: 1.27
Height: 1.00 Width: 6.62 Depth: 8.96
Language: English
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.

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