PMID: 8967436May 1, 1996Paper

Role of myristoylation in membrane attachment and function of G alpha i-3 on Golgi membranes

The American Journal of Physiology
S H BrandJ L Stow

Abstract

Heterotrimeric G protein alpha-subunits localized on the cytoplasmic face of Golgi membranes are involved in regulating vesicle trafficking and protein secretion. We investigated the role of myristoylation in attachment of the G alpha i-3 subunit to Golgi membranes. G alpha i-3 was epitope-tagged by insertion of a FLAG sequence at an NH2-terminal site predicted to interfere with myristoylation, and the resulting NT-alpha i-3 construct was stably transfected and expressed in polarized epithelial LLC-PK1 cells. Metabolic labeling confirmed that the translation product of NT-alpha i-3 was not myristoylated. In contrast to endogenous G alpha 1-3, which is tightly bound to Golgi membranes, the unmyristoylated FLAG-tagged NT-alpha i-3 did not attach to membranes; it was localized by immunofluorescence in the cytoplasm of LLC-PK1 cells and was detected only in the cytosol fraction of cell homogenates. Pertussis toxin-dependent ADP-ribosylation was used to test the ability of NT-alpha i-3 to interact with membrane-bound beta gamma-subunits. In both in vitro and in vivo assays, cytosolic NT-alpha i-3 alone was not ADP-ribosylated, although in the presence of membranes it could interact with G beta gamma-subunits to form heterotrimers. T...Continue Reading

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Citations

Jun 3, 2004·The Journal of Biological Chemistry·Zhixian ZhangTheodore G Wensel
Jul 10, 2003·Annals of the New York Academy of Sciences·R A SturmN G Martin
Mar 27, 2003·Traffic·Fiona G WylieJennifer L Stow
Aug 26, 1998·Biochimica Et Biophysica Acta·J L Stow, K Heimann
Jul 9, 1999·Progress in Lipid Research·J C DeMarR E Anderson
Jun 10, 1998·Microbiology and Molecular Biology Reviews : MMBR·P StanleyC Hughes
Dec 5, 2006·Food and Chemical Toxicology : an International Journal Published for the British Industrial Biological Research Association·George A Burdock, Ioana G Carabin
Sep 13, 2011·Cellular Signalling·Thamara Hewavitharana, Philip B Wedegaertner

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