Description
Humana Press Inc. Two-Hybrid Systems: Methods And Protocols by MACDONALD N.
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.show more