By Hydar Ali and Bodduluri Haribabu
This totally up to date and accelerated moment version of Transmembrane Signaling Protocols demonstrates using such strategies as unmarried phone imaging, bioluminecence resonance strength move, and worldwide proteomics within the research of transmembrane signaling occasions. Highlights contain the practical expression and genetic number of mutant mammalian transmembrane receptors in yeast, real-time research of transmembrane receptor signaling in stay cells, and bioluminescence strength move to watch protein-protein interplay. extra chapters use state of the art easy methods to learn gene expression and proteomic profiles with regards to transmembrane signaling occasions, in addition to to genetically reconstitute bone marrow for the examine of sign transduction ex vivo. The protocols stick to the winning equipment in Molecular Biology™ sequence structure, each one supplying step by step laboratory directions, an creation outlining the rules in the back of the process, lists of the mandatory apparatus and reagents, and pointers on troubleshooting and keeping off recognized pitfalls.
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Additional resources for Transmembrane Signaling Protocols, 2nd Edition (Methods in Molecular Biology Vol 332)
PSD-95 binds to the β1 adrenergic receptor C-terminus and decreases receptor endocytosis (123), whereas another G Protein-Coupled Receptors 25 PDZ domain-containing protein that binds the same C-terminal epitope, membrane-associated guanylate kinase inverted-2 (MAGI-2), a multidomain scaffolding protein containing six PDZ domains, markedly enhances β1 receptor internalization (124). The PDZ domain of the src homology (SH)3 multiple ankrin domain-containing protein (Shank)/somatostatin receptor interacting protein (SSTRIP) associates with the C-terminus of the somatostatin SST2 receptor.
Dale, L. , Anborgh, P. , O’Connor-Halligan, K. , and Ferguson, S. S. (2004) G Protein-coupled receptor kinase 2 RGS homology domain binds to both metabotropic glutamate receptor 1a and G alpha q to attenuate signaling. J. Biol. Chem. 279, 16,614–16,620. 65. Barak, L. , Ferguson, S. , and Caron, M. G. (1997) A beta-arrestin/ green fluorescent protein biosensor for detecting G protein-coupled receptor activation. J. Biol. Chem. 272, 27,497–27,500. 40 Luttrell 66. Oakley, R. , Laporte, S. , Holt, J.
PDZ domaincontaining proteins often possess other protein interaction domains, and commonly serve as scaffolds for protein localization, trafficking, or complex assembly. The C-terminal tails of several GPCRs, including the β2 adrenergic receptor, 5HT2a-c serotonin receptors, SSTR2 somatostatin receptor, the mGluR1a and mGluR5 metabotropic glutamate receptors, and the parathyroid hormone PTH/ PTHrP receptor, end in a PDZ domain-binding motif. Not surprisingly, a number of PDZ domain-containing proteins have been shown to interact with these GPCRs and to play important roles in modifying receptor function (118,119).