G.ALPHA.i3 in Pancreatic Zymogen Granules Participates in Vesicular Fusion.

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概要

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• Earlier studies indicated that a Gi-like protein localized in pancreatic zymogen granule (ZG) membrane mediates vesicle swelling, and is a potentially important prerequisite for vesicle fusion at the cell plasma membrane (PM) [Jena et al. (1997) Proc. Natl. Acad. Sci. USA 94, 13317-13322]. In the present study, we demonstrate the presence of Gαi3 immunoreactivity in ZGs of rat exocrine pancreas using immunoblot assays, light and electron immunomicroscopy. Since GTP has been implicated in the fusion of isolated ZG with PM fractions [Nadin et al. (1989) J. Cell Biol. 109, 2801-2808], the potential role of ZG-associated Gαi3 was investigated. Immunoblot assays demonstrate an increase in Gαi3 protein in ZGs isolated from carbamylcholine stimulated pancreas. Thin layer chromatography shows an increase in GTP hydrolysis by GTPase in ZGs isolated from stimulated compared to resting pancreas. In vitro fusion assays demonstrate that ZGs isolated from carbamylcholine-stimulated pancreatic lobules fuse with the PM at a greater potency in the presence of GTP, mastoparan (G protein agonist) and its analogue mas7. Furthermore, Gαi3-specific a recombinant GAIP (G alpha interacting protein), potenti-ates ZG-PM fusion in the presence of GTP but not in presence of the non-hydrolyzable GTP analogue Gpp(NH)p. Our immunoblot analysis demonstrates the recruitment of Gαi3 immunoreactivity to ZG from stimulated acinar cells, and these isolated ZGs are more potent and efficient in fusing with plasma membrane fractions, suggesting the possible involvement of Gαi3 in ZG-PM fusion. The participation of ZG-associated Gαi3 in ZG-PM fusion is further confirmed by the influence of the Gαi3-specific GAIP, which is known to interact specifically with Gαi3, and not with Gαi2 or Gαq [DeVries et al. (1995) Proc. Natl. Acad. Sci. USA 92, 11916-11920]. Additionally, our data suggest that GTP hydrolysis is a requirement for ZG-PM fusion since GAIP in the presence of Gpp (NH) p shows little or no effect on fusion, whereas GAIP in the presence of GTP significantly potentiates ZG-PM fusion. Our studies suggest a possible role for ZG-associated Gαi3 in ZG-PM fusion.

• 社団法人 日本生化学会の論文

著者

• Jena Bhanu

  Department Of Physiology Wayne State University School Of Medicine

• SATTAR Akm

  Department of Physiology Wayne State University School of Medicine

• BOINPALLY Ramesh

  Department of Physiology Wayne State University School of Medicine

• Stromer Marvin

  Department Of Animal Science And Biochermistry Iowa State University

関連論文

• Impaired Hepatocyte Glucose Transport Protein (GLUT2) Internalization in Chronic Pancreatitis
• Gα_ in Pancreatic Zymogen Granules Participates in Vesicular Fusion
• G.ALPHA.i3 in Pancreatic Zymogen Granules Participates in Vesicular Fusion.

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