A Putative Heterotrimeric G Protein Inhibits the Fusion of COPI-coated Vesicles

• Published in: The Journal of biological chemistry Vol. 273; no. 24; pp. 15203 - 15208
• Main Authors: Helms, J. Bernd, Helms-Brons, Désiré, Brügger, Britta, Gkantiragas, Ioannis, Eberle, Heike, Nickel, Walter, Nürnberg, Bernd, Gerdes, Hans-Hermann, Wieland, Felix T.
• Format: Journal Article
• Language: English
• Published: Elsevier Inc 12-06-1998; American Society for Biochemistry and Molecular Biology
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.273.24.15203

Heterotrimeric G proteins have been implicated in the regulation of intracellular protein transport, but their mechanism of action remains unclear. In vivo , secretion of chromogranin B, tagged with the green fluorescent protein, was inhibited by the addition of a general activator of trimeric G proteins (AlF4−) to stably transfected Vero cells and resulted in an accumulation of the tagged protein in the Golgi apparatus. In an in vitro assay that reconstitutes intra-Golgi protein transport, we find that a membrane-bound and AlF4−-sensitive factor is involved in the fusion reaction. To determine whether this effect is mediated by a heterotrimeric G protein localized to COPI-coated transport vesicles, we determined the presence of G proteins on these vesicles and found that they were segregated relative to the donor membranes. Because G proteins do not have an obvious sorting, retention, or retrieval signal, we considered the possibility that other interactions might be responsible for this segregation. In agreement with this, we found that trimeric G proteins from isolated Golgi membranes were partially insoluble in Triton X-100. Identification of the proteins that interact with the heterotrimeric G proteins in the Golgi-derived detergent-insoluble complex might help to reveal the regulation of protein secretion mediated by heterotrimeric G proteins.