The R-SNARE endobrevin/VAMP-8 mediates homotypic fusion of early endosomes and late endosomes
Endobrevin/VAMP-8 is an R-SNARE localized to endosomes, but it is unknown in which intracellular fusion step it operates. Using subcellular fractionation and quantitative immunogold electron microscopy, we found that endobrevin/VAMP-8 is present on all membranes known to communicate with early endos...
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| Veröffentlicht in: | Molecular biology of the cell 2000-10, Vol.11 (10), p.3289-3298 |
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| creator | Antonin, W Holroyd, C Tikkanen, R Höning, S Jahn, R |
| description | Endobrevin/VAMP-8 is an R-SNARE localized to endosomes, but it is unknown in which intracellular fusion step it operates. Using subcellular fractionation and quantitative immunogold electron microscopy, we found that endobrevin/VAMP-8 is present on all membranes known to communicate with early endosomes, including the plasma membrane, clathrin-coated pits, late endosomes, and membranes of the trans-Golgi network. Affinity-purified antibodies that block the ability of endobrevin/VAMP-8 to form SNARE core complexes potently inhibit homotypic fusion of both early and late endosomes in vitro. Fab fragments were as active as intact immunoglobulin Gs. Recombinant endobrevin/VAMP-8 inhibited both fusion reactions with similar potency. We conclude that endobrevin/VAMP-8 operates as an R-SNARE in the homotypic fusion of early and late endosomes. |
| doi_str_mv | 10.1091/mbc.11.10.3289 |
| format | Article |
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Using subcellular fractionation and quantitative immunogold electron microscopy, we found that endobrevin/VAMP-8 is present on all membranes known to communicate with early endosomes, including the plasma membrane, clathrin-coated pits, late endosomes, and membranes of the trans-Golgi network. Affinity-purified antibodies that block the ability of endobrevin/VAMP-8 to form SNARE core complexes potently inhibit homotypic fusion of both early and late endosomes in vitro. Fab fragments were as active as intact immunoglobulin Gs. Recombinant endobrevin/VAMP-8 inhibited both fusion reactions with similar potency. 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We conclude that endobrevin/VAMP-8 operates as an R-SNARE in the homotypic fusion of early and late endosomes.</description><subject>Animals</subject><subject>Cell Fractionation</subject><subject>Cell Membrane - physiology</subject><subject>Cell Membrane - ultrastructure</subject><subject>Cell-Free System</subject><subject>Coated Pits, Cell-Membrane - physiology</subject><subject>Coated Pits, Cell-Membrane - ultrastructure</subject><subject>Endosomes - physiology</subject><subject>Endosomes - ultrastructure</subject><subject>Membrane Fusion - physiology</subject><subject>Membrane Proteins - genetics</subject><subject>Membrane Proteins - physiology</subject><subject>Qa-SNARE Proteins</subject><subject>R-SNARE Proteins</subject><subject>Rats</subject><subject>SNARE Proteins</subject><subject>Synaptophysin - physiology</subject><subject>trans-Golgi Network - physiology</subject><subject>trans-Golgi Network - ultrastructure</subject><subject>Vesicular Transport Proteins</subject><issn>1059-1524</issn><issn>1939-4586</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkUlPwzAQhS0EYilcOaKcuKV4vDWWuFSoLBKbynJDlh3bNCiJS5wi9d_jQsVymu17MyM9hA4BDwFLOGlMOQRI-ZCSQm6gXZBU5owXYjPlmMscOGE7aC_GN4yBMTHaRjsAmEhMxS56eZy5bJo_3I6nk8y1NpjOfVTtyfP45j4vssbZSvcuZrPQhH45r8rML2IV2iz4zOmuXn6JYmgSo1ub1Yn-be2jLa_r6A7WcYCeziePZ5f59d3F1dn4Oi8Zx33uLWHMWO8sAWGsASYk94IbX4DxIAsiwBo5soZS7kwBThai1NxJ67kQlg7Q6ffe-cKkl0vX9p2u1byrGt0tVdCV-j9pq5l6DR8KmJQkyY_X8i68L1zsVVPF0tW1bl1YRDUilHBW0AQOv8GyCzF2zv-cAKxWfqjkhwJYlSs_kuDo72O_-NoA-gnl_oiZ</recordid><startdate>20001001</startdate><enddate>20001001</enddate><creator>Antonin, W</creator><creator>Holroyd, C</creator><creator>Tikkanen, R</creator><creator>Höning, S</creator><creator>Jahn, R</creator><general>The American Society for Cell Biology</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20001001</creationdate><title>The R-SNARE endobrevin/VAMP-8 mediates homotypic fusion of early endosomes and late endosomes</title><author>Antonin, W ; Holroyd, C ; Tikkanen, R ; Höning, S ; Jahn, R</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c450t-fd244bdfed216bdb14695f65bf81bf198261db97db335eb81e986ca5e9df566d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>Animals</topic><topic>Cell Fractionation</topic><topic>Cell Membrane - physiology</topic><topic>Cell Membrane - ultrastructure</topic><topic>Cell-Free System</topic><topic>Coated Pits, Cell-Membrane - physiology</topic><topic>Coated Pits, Cell-Membrane - ultrastructure</topic><topic>Endosomes - physiology</topic><topic>Endosomes - ultrastructure</topic><topic>Membrane Fusion - physiology</topic><topic>Membrane Proteins - genetics</topic><topic>Membrane Proteins - physiology</topic><topic>Qa-SNARE Proteins</topic><topic>R-SNARE Proteins</topic><topic>Rats</topic><topic>SNARE Proteins</topic><topic>Synaptophysin - physiology</topic><topic>trans-Golgi Network - physiology</topic><topic>trans-Golgi Network - ultrastructure</topic><topic>Vesicular Transport Proteins</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Antonin, W</creatorcontrib><creatorcontrib>Holroyd, C</creatorcontrib><creatorcontrib>Tikkanen, R</creatorcontrib><creatorcontrib>Höning, S</creatorcontrib><creatorcontrib>Jahn, R</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Molecular biology of the cell</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Antonin, W</au><au>Holroyd, C</au><au>Tikkanen, R</au><au>Höning, S</au><au>Jahn, R</au><au>Pfeffer, Suzanne R.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The R-SNARE endobrevin/VAMP-8 mediates homotypic fusion of early endosomes and late endosomes</atitle><jtitle>Molecular biology of the cell</jtitle><addtitle>Mol Biol Cell</addtitle><date>2000-10-01</date><risdate>2000</risdate><volume>11</volume><issue>10</issue><spage>3289</spage><epage>3298</epage><pages>3289-3298</pages><issn>1059-1524</issn><eissn>1939-4586</eissn><abstract>Endobrevin/VAMP-8 is an R-SNARE localized to endosomes, but it is unknown in which intracellular fusion step it operates. 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| subjects | Animals Cell Fractionation Cell Membrane - physiology Cell Membrane - ultrastructure Cell-Free System Coated Pits, Cell-Membrane - physiology Coated Pits, Cell-Membrane - ultrastructure Endosomes - physiology Endosomes - ultrastructure Membrane Fusion - physiology Membrane Proteins - genetics Membrane Proteins - physiology Qa-SNARE Proteins R-SNARE Proteins Rats SNARE Proteins Synaptophysin - physiology trans-Golgi Network - physiology trans-Golgi Network - ultrastructure Vesicular Transport Proteins |
| title | The R-SNARE endobrevin/VAMP-8 mediates homotypic fusion of early endosomes and late endosomes |
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