caa a22 4500 606236953 CHVBK 20210128101250.0 cr unu---uuuuu 210128e20150301xx s 000 0 eng 10.1007/s11010-014-2289-7 doi (NATIONALLICENCE)springer-10.1007/s11010-014-2289-7 Activator of G protein signaling 3 forms a complex with resistance to inhibitors of cholinesterase-8A without promoting nucleotide exchange on Gαi3 [Elektronische Daten] [Man Tse, Christina Morris, Mingjie Zhang, Yung Wong] Activator of G protein signaling 3 (AGS3) is a guanine nucleotide dissociation inhibitor (GDI) which stabilizes the Gαi/o subunits as an AGS3/Gαi/o-GDP complex. It has recently been demonstrated in reconstitution experiments that the AGS3/Gαi/o-GDP complex may act as a substrate of resistance to inhibitors of cholinesterase 8A (Ric-8A), a guanine exchange factor (GEF) for heterotrimeric Gα proteins. Since the ability of Ric-8A to activate Gαi/o subunits that are bound to AGS3 in a cellular environment has not been confirmed, we thus examined the effect of Ric-8A on cAMP accumulation in HEK293 cells expressing different forms of AGS3 and Gαi3. Co-immunoprecipitation assays indicate that full-length AGS3 and its N- and C-terminal truncated mutants can interact with Ric-8A in HEK293 cells. Yeast two-hybrid assay further confirmed that Ric-8A can directly bind to AGS3S, a short form of AGS3 which is endogenously expressed in heart. However, Ric-8A failed to facilitate Gαi-induced suppression of adenylyl cyclase, suggesting that it may not serve as a GEF for AGS3/Gαi/o-GDP complex in a cellular environment. Springer Science+Business Media New York, 2014 AGS3 nationallicence GPSM1 nationallicence Ric-8A nationallicence GEF nationallicence cAMP nationallicence Tse Man Division of Life Science and the Biotechnology Research Institute, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China aut Morris Christina Division of Life Science and the Biotechnology Research Institute, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China aut Zhang Mingjie Division of Life Science and the Biotechnology Research Institute, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China aut Wong Yung Division of Life Science and the Biotechnology Research Institute, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China aut Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 401/1-2(2015-03-01), 27-38 0300-8177 401:1-2<27 2015 401 11010 https://doi.org/10.1007/s11010-014-2289-7 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s11010-014-2289-7 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Tse Man Division of Life Science and the Biotechnology Research Institute, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China aut NATIONALLICENCE 700 1- Morris Christina Division of Life Science and the Biotechnology Research Institute, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China aut NATIONALLICENCE 700 1- Zhang Mingjie Division of Life Science and the Biotechnology Research Institute, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China aut NATIONALLICENCE 700 1- Wong Yung Division of Life Science and the Biotechnology Research Institute, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China aut NATIONALLICENCE 773 0- Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 401/1-2(2015-03-01), 27-38 0300-8177 401:1-2<27 2015 401 11010