Ca2+-independent,protein-mediated fusion of chromaffin granule ghosts with liposomes |
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Authors: | Michal Bental Peter Lelkes Janny Scholma Dick Hoekstra Jan Wilschut |
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Affiliation: | 1. Department of Membrane Research, The Weizmann Institute of Science, P.O.B. 26, Rehovot 76100 Israel;2. Laboratory of Physiological Chemistry, University of Groningen, Bloemsingel 10, 9712 KZ Groningen Netherlands |
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Abstract: | We have investigated the interaction between isolated membrane vesicles from chromaffin granules and large unilamellar phospholipid vesicles (liposomes). Mixing of membrane lipids has been monitored continuously, utilizing the fluorescence resonance energy transfer assay described by Struck et al. ((1982) Biochemistry 20, 4093–4099). To demonstrate coalescence of the internal vesicle volumes the transfer of colloidal gold from the liposomes to the interior of the granule membrane vesicles has been examined. Efficient fusion of the liposomes with the granule membranes was observed. Significant fusion occurred in the absence of Ca2+, although the extent of interaction was enhanced in its presence. The sensitivity of the interaction to pretreatment of the granule membranes with trypsin showed the fusion reaction to be a protein-mediated process. |
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Keywords: | Chromaffin granule Exocytosis Liposome Phospholipid vesicle Membrane fusion Chol cholesterol CL cardiolipin EGTA Hepes LUV large unilamellar vesicles PC phosphatidylcholine RET resonance energy transfer |
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