Reconstitution of calcium-triggered membrane fusion using "reserve" granules.

The Journal of Biological Chemistry
V V ChestkovS S Vogel

Abstract

Calcium-gated secretion of proteins involves the transfer of "reserve" granules, exocytotic vesicles that are cytoplasmic and, hence, plasma membrane-naive, from the cell interior to the surface membrane where they dock prior to fusion. Docking and subsequent priming steps are thought to require cytoplasmic factors. These steps are believed to induce fusion competence. We have tested this hypothesis by isolating reserve granules from sea urchin eggs and determining under which conditions these granules will fuse. We find that isolated reserve granules, lacking soluble cofactors, support calcium-dependent membrane fusion in vitro. Preincubation with adenosine 5'-3-O-(thio)triphosphate and guanosine 5'-3-O-(thio)triphosphate did not prevent fusion. Thus, isolated reserve granules have all the necessary components required for calcium-gated fusion prior to docking.

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Citations

Jun 19, 2003·Developmental Biology·Wojciech FrejtagSteven S Vogel
Dec 22, 1999·Pharmaceutical Science & Technology Today·C R Lewanski, S Stewart
Jun 2, 2005·Nature Reviews. Molecular Cell Biology·Paul L McNeil, Tom Kirchhausen
Jan 25, 2012·PLoS Biology·Kannan AlpadiChristopher Peters
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Mar 7, 2001·Nature Cell Biology·T NemotoH Kasai
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