Covalent Cross-Linking of the Phytophthora megasperma Oligopeptide Elicitor to its Receptor in Parsley Membranes
An oligopeptide elicitor from Phytophthora megasperma f.sp. glycinea (Pep-13) that induces phytoalexin accumulation in cultured parsley cells was radioiodinated and chemically cross-linked to its binding site in microsomal and plasma membrane preparations with each of three homobifunctional reagents...
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Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 1995-03, Vol.92 (6), p.2338-2342 |
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description | An oligopeptide elicitor from Phytophthora megasperma f.sp. glycinea (Pep-13) that induces phytoalexin accumulation in cultured parsley cells was radioiodinated and chemically cross-linked to its binding site in microsomal and plasma membrane preparations with each of three homobifunctional reagents. Analysis by SDS/PAGE and autoradiography of solubilized membrane proteins demonstrated labeling of a 91-kDa protein, regardless of which reagent was used. Cross-linking of this protein was prevented by addition of excess unlabeled Pep-13. The competitor concentration found to half-maximally reduce the intensity of the cross-linked band was 6 nM, which is in good agreement with the IC50value of 4.7 nM, obtained from ligand binding assays. No cross-linking of125I-labeled Pep-13 was observed by using microsomal membranes from three other plant species, indicating species-specific occurrence of the binding site. Coupling of125I-Pep-13 to the parsley 91-kDa protein required the same structural elements within the ligand as was recently reported for binding of125I-Pep-13 to parsley microsomes, elicitor-induced stimulation of ion fluxes across the plasma membrane, the oxidative burst, the expression of defense-related genes, and phytoalexin production. These findings suggest that the 91-kDa protein identified in parsley membranes is the oligopeptide elicitor receptor mediating activation of a multicomponent defense response. |
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Analysis by SDS/PAGE and autoradiography of solubilized membrane proteins demonstrated labeling of a 91-kDa protein, regardless of which reagent was used. Cross-linking of this protein was prevented by addition of excess unlabeled Pep-13. The competitor concentration found to half-maximally reduce the intensity of the cross-linked band was 6 nM, which is in good agreement with the IC50value of 4.7 nM, obtained from ligand binding assays. No cross-linking of125I-labeled Pep-13 was observed by using microsomal membranes from three other plant species, indicating species-specific occurrence of the binding site. Coupling of125I-Pep-13 to the parsley 91-kDa protein required the same structural elements within the ligand as was recently reported for binding of125I-Pep-13 to parsley microsomes, elicitor-induced stimulation of ion fluxes across the plasma membrane, the oxidative burst, the expression of defense-related genes, and phytoalexin production. These findings suggest that the 91-kDa protein identified in parsley membranes is the oligopeptide elicitor receptor mediating activation of a multicomponent defense response.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.92.6.2338</identifier><identifier>PMID: 7892267</identifier><language>eng</language><publisher>United States: National Academy of Sciences of the United States of America</publisher><subject>Amino Acid Sequence ; Binding sites ; Binding, Competitive ; Cell membranes ; Cells, Cultured ; Cross-Linking Reagents - pharmacology ; Cultured cells ; Daucus carota - metabolism ; Electrophoresis, Polyacrylamide Gel ; Fungal Proteins - isolation & purification ; Fungal Proteins - metabolism ; Glycine max - metabolism ; Glycoproteins ; Intracellular Membranes - metabolism ; Iodine Radioisotopes ; Kinetics ; Ligands ; Magnoliopsida - metabolism ; Membrane Glycoproteins - isolation & purification ; Membrane Glycoproteins - metabolism ; Membrane Proteins ; Microsomes ; Microsomes - metabolism ; Molecular Sequence Data ; P branes ; Parsley ; Peptide Fragments - chemistry ; Peptide Fragments - metabolism ; Peptide Fragments - pharmacology ; Phytoalexins ; Phytophthora - metabolism ; Phytophthora megasperma ; Plant Extracts - biosynthesis ; Plants ; Reagents ; Receptors ; Sesquiterpenes ; Substrate Specificity ; Succinimides - pharmacology ; Terpenes</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 1995-03, Vol.92 (6), p.2338-2342</ispartof><rights>Copyright 1995 The National Academy of Sciences of the United States of America</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c554t-1f0c61fc8689c7130d5de71673acbc3023d90cc74ca692395904b7c4cd896edc3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/92/6.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/2366988$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/2366988$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,723,776,780,799,881,27903,27904,53769,53771,57995,58228</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/7892267$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Nurnberger, Thorsten</creatorcontrib><creatorcontrib>Nennstiel, Dirk</creatorcontrib><creatorcontrib>Hahlbrock, Klaus</creatorcontrib><creatorcontrib>Scheel, Dierk</creatorcontrib><title>Covalent Cross-Linking of the Phytophthora megasperma Oligopeptide Elicitor to its Receptor in Parsley Membranes</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>An oligopeptide elicitor from Phytophthora megasperma f.sp. glycinea (Pep-13) that induces phytoalexin accumulation in cultured parsley cells was radioiodinated and chemically cross-linked to its binding site in microsomal and plasma membrane preparations with each of three homobifunctional reagents. Analysis by SDS/PAGE and autoradiography of solubilized membrane proteins demonstrated labeling of a 91-kDa protein, regardless of which reagent was used. Cross-linking of this protein was prevented by addition of excess unlabeled Pep-13. The competitor concentration found to half-maximally reduce the intensity of the cross-linked band was 6 nM, which is in good agreement with the IC50value of 4.7 nM, obtained from ligand binding assays. No cross-linking of125I-labeled Pep-13 was observed by using microsomal membranes from three other plant species, indicating species-specific occurrence of the binding site. Coupling of125I-Pep-13 to the parsley 91-kDa protein required the same structural elements within the ligand as was recently reported for binding of125I-Pep-13 to parsley microsomes, elicitor-induced stimulation of ion fluxes across the plasma membrane, the oxidative burst, the expression of defense-related genes, and phytoalexin production. These findings suggest that the 91-kDa protein identified in parsley membranes is the oligopeptide elicitor receptor mediating activation of a multicomponent defense response.</description><subject>Amino Acid Sequence</subject><subject>Binding sites</subject><subject>Binding, Competitive</subject><subject>Cell membranes</subject><subject>Cells, Cultured</subject><subject>Cross-Linking Reagents - pharmacology</subject><subject>Cultured cells</subject><subject>Daucus carota - metabolism</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Fungal Proteins - isolation & purification</subject><subject>Fungal Proteins - metabolism</subject><subject>Glycine max - metabolism</subject><subject>Glycoproteins</subject><subject>Intracellular Membranes - metabolism</subject><subject>Iodine Radioisotopes</subject><subject>Kinetics</subject><subject>Ligands</subject><subject>Magnoliopsida - metabolism</subject><subject>Membrane Glycoproteins - isolation & purification</subject><subject>Membrane Glycoproteins - metabolism</subject><subject>Membrane Proteins</subject><subject>Microsomes</subject><subject>Microsomes - metabolism</subject><subject>Molecular Sequence Data</subject><subject>P branes</subject><subject>Parsley</subject><subject>Peptide Fragments - chemistry</subject><subject>Peptide Fragments - metabolism</subject><subject>Peptide Fragments - pharmacology</subject><subject>Phytoalexins</subject><subject>Phytophthora - metabolism</subject><subject>Phytophthora megasperma</subject><subject>Plant Extracts - biosynthesis</subject><subject>Plants</subject><subject>Reagents</subject><subject>Receptors</subject><subject>Sesquiterpenes</subject><subject>Substrate Specificity</subject><subject>Succinimides - pharmacology</subject><subject>Terpenes</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1995</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkUuLFDEUhYMoYzu6daWQlbsq86jKA9xIMz6gZQbRdUinUl0ZU5UySQ_2vzdFt00LgquQnO_c3HsPAC8xqjHi9O086VRLUrOaUCoegRVGEleskegxWCFEeCUa0jwFz1K6RwjJVqArcMWFJITxFZjX4UF7O2W4jiGlauOmH27awdDDPFh4NxxymIc8hKjhaHc6zTaOGt56twuznbPrLLzxzrgcIswBupzgV2uKUu5ugnc6Jm8P8Isdt1FPNj0HT3rtk31xOq_B9w8339afqs3tx8_r95vKtG2TK9wjw3BvBBPScExR13aWY8apNltDEaGdRMbwxmgmCZWtRM2Wm8Z0QjLbGXoN3h3rzvvtWB7KiFF7NUc36nhQQTv1tzK5Qe3Cgyrb4qLY35zsMfzc25TV6JKx3pcZwj4pzrEggv4fLC0TRlhbwPoImmXR0fbnXjBSS5RqiVJJophaoiyG15cTnPFTdhc_L74_6tmv-r332f7KF4X-CRb91VG_TyW1M0AoY1II-htrgb6D</recordid><startdate>19950314</startdate><enddate>19950314</enddate><creator>Nurnberger, Thorsten</creator><creator>Nennstiel, Dirk</creator><creator>Hahlbrock, Klaus</creator><creator>Scheel, Dierk</creator><general>National Academy of Sciences of the United States of America</general><general>National Acad Sciences</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>M7N</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>19950314</creationdate><title>Covalent Cross-Linking of the Phytophthora megasperma Oligopeptide Elicitor to its Receptor in Parsley Membranes</title><author>Nurnberger, Thorsten ; Nennstiel, Dirk ; Hahlbrock, Klaus ; Scheel, Dierk</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c554t-1f0c61fc8689c7130d5de71673acbc3023d90cc74ca692395904b7c4cd896edc3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1995</creationdate><topic>Amino Acid Sequence</topic><topic>Binding sites</topic><topic>Binding, Competitive</topic><topic>Cell membranes</topic><topic>Cells, Cultured</topic><topic>Cross-Linking Reagents - pharmacology</topic><topic>Cultured cells</topic><topic>Daucus carota - metabolism</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Fungal Proteins - isolation & purification</topic><topic>Fungal Proteins - metabolism</topic><topic>Glycine max - metabolism</topic><topic>Glycoproteins</topic><topic>Intracellular Membranes - metabolism</topic><topic>Iodine Radioisotopes</topic><topic>Kinetics</topic><topic>Ligands</topic><topic>Magnoliopsida - metabolism</topic><topic>Membrane Glycoproteins - isolation & purification</topic><topic>Membrane Glycoproteins - metabolism</topic><topic>Membrane Proteins</topic><topic>Microsomes</topic><topic>Microsomes - metabolism</topic><topic>Molecular Sequence Data</topic><topic>P branes</topic><topic>Parsley</topic><topic>Peptide Fragments - chemistry</topic><topic>Peptide Fragments - metabolism</topic><topic>Peptide Fragments - pharmacology</topic><topic>Phytoalexins</topic><topic>Phytophthora - metabolism</topic><topic>Phytophthora megasperma</topic><topic>Plant Extracts - biosynthesis</topic><topic>Plants</topic><topic>Reagents</topic><topic>Receptors</topic><topic>Sesquiterpenes</topic><topic>Substrate Specificity</topic><topic>Succinimides - pharmacology</topic><topic>Terpenes</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Nurnberger, Thorsten</creatorcontrib><creatorcontrib>Nennstiel, Dirk</creatorcontrib><creatorcontrib>Hahlbrock, Klaus</creatorcontrib><creatorcontrib>Scheel, Dierk</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Nurnberger, Thorsten</au><au>Nennstiel, Dirk</au><au>Hahlbrock, Klaus</au><au>Scheel, Dierk</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Covalent Cross-Linking of the Phytophthora megasperma Oligopeptide Elicitor to its Receptor in Parsley Membranes</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>1995-03-14</date><risdate>1995</risdate><volume>92</volume><issue>6</issue><spage>2338</spage><epage>2342</epage><pages>2338-2342</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><abstract>An oligopeptide elicitor from Phytophthora megasperma f.sp. glycinea (Pep-13) that induces phytoalexin accumulation in cultured parsley cells was radioiodinated and chemically cross-linked to its binding site in microsomal and plasma membrane preparations with each of three homobifunctional reagents. Analysis by SDS/PAGE and autoradiography of solubilized membrane proteins demonstrated labeling of a 91-kDa protein, regardless of which reagent was used. Cross-linking of this protein was prevented by addition of excess unlabeled Pep-13. The competitor concentration found to half-maximally reduce the intensity of the cross-linked band was 6 nM, which is in good agreement with the IC50value of 4.7 nM, obtained from ligand binding assays. No cross-linking of125I-labeled Pep-13 was observed by using microsomal membranes from three other plant species, indicating species-specific occurrence of the binding site. Coupling of125I-Pep-13 to the parsley 91-kDa protein required the same structural elements within the ligand as was recently reported for binding of125I-Pep-13 to parsley microsomes, elicitor-induced stimulation of ion fluxes across the plasma membrane, the oxidative burst, the expression of defense-related genes, and phytoalexin production. These findings suggest that the 91-kDa protein identified in parsley membranes is the oligopeptide elicitor receptor mediating activation of a multicomponent defense response.</abstract><cop>United States</cop><pub>National Academy of Sciences of the United States of America</pub><pmid>7892267</pmid><doi>10.1073/pnas.92.6.2338</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Amino Acid Sequence Binding sites Binding, Competitive Cell membranes Cells, Cultured Cross-Linking Reagents - pharmacology Cultured cells Daucus carota - metabolism Electrophoresis, Polyacrylamide Gel Fungal Proteins - isolation & purification Fungal Proteins - metabolism Glycine max - metabolism Glycoproteins Intracellular Membranes - metabolism Iodine Radioisotopes Kinetics Ligands Magnoliopsida - metabolism Membrane Glycoproteins - isolation & purification Membrane Glycoproteins - metabolism Membrane Proteins Microsomes Microsomes - metabolism Molecular Sequence Data P branes Parsley Peptide Fragments - chemistry Peptide Fragments - metabolism Peptide Fragments - pharmacology Phytoalexins Phytophthora - metabolism Phytophthora megasperma Plant Extracts - biosynthesis Plants Reagents Receptors Sesquiterpenes Substrate Specificity Succinimides - pharmacology Terpenes |
title | Covalent Cross-Linking of the Phytophthora megasperma Oligopeptide Elicitor to its Receptor in Parsley Membranes |
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