title: Biochemical, Biophysical and Cellular Techniques to Study the Guanine Nucleotide Exchange Factor, GIV/Girdin.
abstract: Canonical signal transduction via heterotrimeric G proteins is spatiotemporally restricted, i.e., triggered exclusively at the plasma membrane, only by agonist activation of G protein-coupled receptors via a finite process that is terminated within a few hundred milliseconds. Recently, a rapidly emerging paradigm has revealed a noncanonical pathway for activation of heterotrimeric G proteins via the nonreceptor guanidine-nucleotide exchange factor, GIV/Girdin. Biochemical, biophysical, and functional studies evaluating this pathway have unraveled its unique properties and distinctive spatiotemporal features. As in the case of any new pathway/paradigm, these studies first required an in-depth optimization of tools/techniques and protocols, governed by rationale and fundamentals unique to the pathway, and more specifically to the large multimodular GIV protein. Here we provide the most up-to-date overview of protocols that have generated most of what we know today about noncanonical G protein activation by GIV and its relevance in health and disease. © 2016 by John Wiley & Sons, Inc.
authors.affiliation: Department of Cellular and Molecular Medicine, University of California at San Diego, La Jolla, California.|Department of Medicine, University of California at San Diego, La Jolla, California.
authors.forename: Pradipta
authors.initials: P
authors.lastname: Ghosh
authors.affiliation: Department of Medicine, University of California at San Diego, La Jolla, California.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Aznar
authors.affiliation: Department of Medicine, University of California at San Diego, La Jolla, California.
authors.forename: Lee
authors.initials: L
authors.lastname: Swanson
authors.affiliation: Department of Cellular and Molecular Medicine, University of California at San Diego, La Jolla, California.
authors.forename: I-Chung
authors.initials: IC
authors.lastname: Lo
authors.affiliation: Department of Medicine, University of California at San Diego, La Jolla, California.
authors.forename: Inmaculada
authors.initials: I
authors.lastname: Lopez-Sanchez
authors.affiliation: Department of Medicine, University of California at San Diego, La Jolla, California.
authors.forename: Jason
authors.initials: J
authors.lastname: Ear
authors.affiliation: Department of Medicine, University of California at San Diego, La Jolla, California.
authors.forename: Cristina
authors.initials: C
authors.lastname: Rohena
authors.affiliation: Department of Medicine, University of California at San Diego, La Jolla, California.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Kalogriopoulos
authors.affiliation: Department of Medicine, University of California at San Diego, La Jolla, California.
authors.forename: Linda
authors.initials: L
authors.lastname: Joosen
authors.affiliation: Department of Medicine, University of California at San Diego, La Jolla, California.
authors.forename: Ying
authors.initials: Y
authors.lastname: Dunkel
authors.affiliation: Department of Medicine, University of California at San Diego, La Jolla, California.
authors.forename: Nina
authors.initials: N
authors.lastname: Sun
authors.affiliation: Department of Chemistry and Biochemistry, California State University Long Beach, Long Beach, California.
authors.forename: Peter
authors.initials: P
authors.lastname: Nguyen
authors.affiliation: Department of Chemistry and Biochemistry, California State University Long Beach, Long Beach, California.
authors.forename: Deepali
authors.initials: D
authors.lastname: Bhandari
mesh_terms: D000818:Animals; D001703:Biophysics / Q000379:methods; D005455:Fluorescent Antibody Technique / Q000379:methods*; D020662:Guanine Nucleotide Exchange Factors / Q000032:analysis* / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting / Q000379:methods*; D047468:Immunoprecipitation / Q000379:methods*; D015398:Signal Transduction
chemical_list: D020662:Guanine Nucleotide Exchange Factors
keywords: FRET; GIV/Girdin; immunoblotting; immunofluorescence; immunoprecipitation; in cellulo GST-pull down; trimeric G proteins