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Characterization of Constitutively Active Mutants of G Protein-Coupled Receptors

heterotrimer. Some mutants of G protein-coupled receptors, known as constitutively active mutants, have the capac-ity to activate the G protein-signaling cascade even in the absence of ligand. In this chapter, we describe three methods that most directly allow

Expression and Purification of Active Protein Kinases from Wheat Germ Extracts

In vitro functional studies of eukaryotic kinases are often constrained by the availability of pure and �enzymatically active kinase of interest. Though numerous proteins have been synthesized by cell-based systems, in vivo production

Active Site Profiling to Identify Protein Functional Sites in Sequences and Structures Using the Deacon Active Site Profiler (DASP)

) to allow users to create an active site profile for any protein family, is described. Two protocols for functional site analysis of protein families using DASP are presented: 1) creation of functional site signatures and a profile from proteins of known