Mitochondrial dolichyl-phosphate mannose synthase : purification and immunogold localization by electron microscopy
Mitochondrial dolichyl-phosphate mannose synthase has been purified to homogeneity using an original procedure, reconstitution into specific phospholipid vesicles and sedimentation on a sucrose gradient as final step. The enzyme has an apparent molecular mass of 30 kDa on an SDS/polyacrylamide gel....
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| Veröffentlicht in: | European Journal of Biochemistry 1992-06, Vol.206 (3), p.853-858 |
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| container_issue | 3 |
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| container_title | European Journal of Biochemistry |
| container_volume | 206 |
| creator | GASNIER, F ROUSSON, R LERME, F VAGANAY, E LOUISOT, P GATEAU-ROESCH, O |
| description | Mitochondrial dolichyl-phosphate mannose synthase has been purified to homogeneity using an original procedure, reconstitution into specific phospholipid vesicles and sedimentation on a sucrose gradient as final step. The enzyme has an apparent molecular mass of 30 kDa on an SDS/polyacrylamide gel. Increased enzyme activity could be correlated with this polypeptide band. A specific antibody was raised in rabbits against this transferase. Specific IgG obtained from the immune serum removed enzymatic activity from a detergent extract of mitochondrial outer membrane and reacted specifically with the 30-kDa band on immunoblots. Furthermore, an immunocytochemical experiment proved the localization of dolichyl-phosphate mannose synthase on the cytosolic face of the outer membrane of mitochondria. |
| doi_str_mv | 10.1111/j.1432-1033.1992.tb16993.x |
| format | Article |
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The enzyme has an apparent molecular mass of 30 kDa on an SDS/polyacrylamide gel. Increased enzyme activity could be correlated with this polypeptide band. A specific antibody was raised in rabbits against this transferase. Specific IgG obtained from the immune serum removed enzymatic activity from a detergent extract of mitochondrial outer membrane and reacted specifically with the 30-kDa band on immunoblots. Furthermore, an immunocytochemical experiment proved the localization of dolichyl-phosphate mannose synthase on the cytosolic face of the outer membrane of mitochondria.</description><subject>Analytical, structural and metabolic biochemistry</subject><subject>Animals</subject><subject>Biochemistry, Molecular Biology</subject><subject>Biological and medical sciences</subject><subject>Blotting, Western</subject><subject>Chromatography, DEAE-Cellulose</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Enzymes and enzyme inhibitors</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Immunoglobulin G</subject><subject>Immunohistochemistry</subject><subject>Intracellular Membranes - enzymology</subject><subject>Life Sciences</subject><subject>Mannosyltransferases - analysis</subject><subject>Mannosyltransferases - antagonists & inhibitors</subject><subject>Mannosyltransferases - isolation & purification</subject><subject>Mice</subject><subject>Microscopy, Electron</subject><subject>Mitochondria, Liver - enzymology</subject><subject>Molecular Weight</subject><subject>Transferases</subject><issn>0014-2956</issn><issn>1432-1033</issn><issn>1432-1327</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1992</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpFkUtv1DAUhS0EKtPCT0CKEELqIsGPxGN3V1XQVhrEBtaWXyEeOXawE9Tw60ma0dQbH_mce617PwA-Ilih5Xw5VqgmuESQkApxjqtRIco5qZ5egd3Zeg12EKK6xLyhb8FlzkcIIeV0fwEuEH1WO5C_uzHqLgaTnPSFid7pbvbl0MU8dHK0RS9DiNkWeQ5jJxdxUwxTcq3TcnQxFDKYwvX9FOLv6E3ho5be_ds8NRfWWz2mRfdOp5h1HOZ34E0rfbbvT_cV-PXt68-7h_Lw4_7x7vZQaoLhWNYUE25ojfawoVDCWhmiGWe4YZhpY1plIJbIYIQbpYxUkBnJW0sUQ7TWmFyB661vJ70YkutlmkWUTjzcHsT6BiFBdU3QX7RkP2_ZIcU_k82j6F3W1nsZbJyy2GPOKWZsCd5swXWanGx77oygWOmIo1gRiBWBWOmIEx3xtBR_OP0yqd6al9INx-J_OvkyL2tskwza5XOsIYzsGST_AcQ8m4Q</recordid><startdate>19920615</startdate><enddate>19920615</enddate><creator>GASNIER, F</creator><creator>ROUSSON, R</creator><creator>LERME, F</creator><creator>VAGANAY, E</creator><creator>LOUISOT, P</creator><creator>GATEAU-ROESCH, O</creator><general>Blackwell</general><general>Wiley</general><scope>IQODW</scope><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>1XC</scope></search><sort><creationdate>19920615</creationdate><title>Mitochondrial dolichyl-phosphate mannose synthase : purification and immunogold localization by electron microscopy</title><author>GASNIER, F ; ROUSSON, R ; LERME, F ; VAGANAY, E ; LOUISOT, P ; GATEAU-ROESCH, O</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c320t-46239d64170560a04bd3c89825828cddfbd02a1d2125bbdab08da9fe3b8164c23</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1992</creationdate><topic>Analytical, structural and metabolic biochemistry</topic><topic>Animals</topic><topic>Biochemistry, Molecular Biology</topic><topic>Biological and medical sciences</topic><topic>Blotting, Western</topic><topic>Chromatography, DEAE-Cellulose</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Enzymes and enzyme inhibitors</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Immunoglobulin G</topic><topic>Immunohistochemistry</topic><topic>Intracellular Membranes - enzymology</topic><topic>Life Sciences</topic><topic>Mannosyltransferases - analysis</topic><topic>Mannosyltransferases - antagonists & inhibitors</topic><topic>Mannosyltransferases - isolation & purification</topic><topic>Mice</topic><topic>Microscopy, Electron</topic><topic>Mitochondria, Liver - enzymology</topic><topic>Molecular Weight</topic><topic>Transferases</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>GASNIER, F</creatorcontrib><creatorcontrib>ROUSSON, R</creatorcontrib><creatorcontrib>LERME, F</creatorcontrib><creatorcontrib>VAGANAY, E</creatorcontrib><creatorcontrib>LOUISOT, P</creatorcontrib><creatorcontrib>GATEAU-ROESCH, O</creatorcontrib><collection>Pascal-Francis</collection><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>Hyper Article en Ligne (HAL)</collection><jtitle>European Journal of Biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>GASNIER, F</au><au>ROUSSON, R</au><au>LERME, F</au><au>VAGANAY, E</au><au>LOUISOT, P</au><au>GATEAU-ROESCH, O</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Mitochondrial dolichyl-phosphate mannose synthase : purification and immunogold localization by electron microscopy</atitle><jtitle>European Journal of Biochemistry</jtitle><addtitle>Eur J Biochem</addtitle><date>1992-06-15</date><risdate>1992</risdate><volume>206</volume><issue>3</issue><spage>853</spage><epage>858</epage><pages>853-858</pages><issn>0014-2956</issn><eissn>1432-1033</eissn><eissn>1432-1327</eissn><coden>EJBCAI</coden><abstract>Mitochondrial dolichyl-phosphate mannose synthase has been purified to homogeneity using an original procedure, reconstitution into specific phospholipid vesicles and sedimentation on a sucrose gradient as final step. The enzyme has an apparent molecular mass of 30 kDa on an SDS/polyacrylamide gel. Increased enzyme activity could be correlated with this polypeptide band. A specific antibody was raised in rabbits against this transferase. Specific IgG obtained from the immune serum removed enzymatic activity from a detergent extract of mitochondrial outer membrane and reacted specifically with the 30-kDa band on immunoblots. Furthermore, an immunocytochemical experiment proved the localization of dolichyl-phosphate mannose synthase on the cytosolic face of the outer membrane of mitochondria.</abstract><cop>Oxford</cop><pub>Blackwell</pub><pmid>1606967</pmid><doi>10.1111/j.1432-1033.1992.tb16993.x</doi><tpages>6</tpages></addata></record> |
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| identifier | ISSN: 0014-2956 |
| ispartof | European Journal of Biochemistry, 1992-06, Vol.206 (3), p.853-858 |
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| language | eng |
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| source | MEDLINE; Alma/SFX Local Collection |
| subjects | Analytical, structural and metabolic biochemistry Animals Biochemistry, Molecular Biology Biological and medical sciences Blotting, Western Chromatography, DEAE-Cellulose Electrophoresis, Polyacrylamide Gel Enzymes and enzyme inhibitors Fundamental and applied biological sciences. Psychology Immunoglobulin G Immunohistochemistry Intracellular Membranes - enzymology Life Sciences Mannosyltransferases - analysis Mannosyltransferases - antagonists & inhibitors Mannosyltransferases - isolation & purification Mice Microscopy, Electron Mitochondria, Liver - enzymology Molecular Weight Transferases |
| title | Mitochondrial dolichyl-phosphate mannose synthase : purification and immunogold localization by electron microscopy |
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