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dc.contributor.authorKellaris, Kennan V.
dc.contributor.authorBowen, Sharon
dc.contributor.authorGilmore, Reid
dc.date2022-08-11T08:10:05.000
dc.date.accessioned2022-08-23T16:55:03Z
dc.date.available2022-08-23T16:55:03Z
dc.date.issued1991-07-01
dc.date.submitted2008-10-31
dc.identifier.citationJ Cell Biol. 1991 Jul;114(1):21-33.
dc.identifier.issn0021-9525 (Print)
dc.identifier.pmid1646822
dc.identifier.urihttp://hdl.handle.net/20.500.14038/42634
dc.description.abstractWe have used the homobifunctional cross-linking reagent disuccinimidyl suberate (DSS) to identify proteins that are adjacent to nascent polypeptides undergoing translocations across mammalian rough ER. Translocation intermediates were assembled by supplementing cell free translations of truncated mRNAs with the signal recognition particle (SRP) and microsomal membrane vesicles. Two prominent cross-linked products of 45 and 64 kD were detected. The 64-kD product was obtained when the cell free translation contained SRP, while formation of the 45-kD product required both SRP and translocation competent microsomal membrane vesicles. In agreement with previous investigators, we suggest that the 64-kD product arises by cross-linking of the nascent polypeptide to the 54-kD subunit of SRP. The 45-kD product resists alkaline extraction from the membrane, so we conclude that the 11-kD nascent polypeptide has been crosslinked to an integral membrane protein of approximately 34 kD (imp34). The cross-linked product does not bind to ConA Sepharose, nor is it sensitive to endoglycosidase H digestion; hence imp34 is not identical to the alpha or beta subunits of the signal sequence receptor (SSR). We propose that imp34 functions in concert with SSR to form a translocation site through which nascent polypeptides pass in traversing the membrane bilayer of the rough endoplasmic reticulum.
dc.language.isoen_US
dc.relation<a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=1646822&dopt=Abstract">Link to Article in PubMed</a>
dc.subject*Calcium-Binding Proteins
dc.subjectCell-Free System
dc.subjectCross-Linking Reagents
dc.subjectEndoplasmic Reticulum
dc.subjectEthylmaleimide
dc.subject*Membrane Glycoproteins
dc.subjectMembrane Proteins
dc.subjectPeptides
dc.subjectRNA, Transfer
dc.subjectRNA, Transfer, Amino Acyl
dc.subjectReceptors, Cell Surface
dc.subject*Receptors, Cytoplasmic and Nuclear
dc.subject*Receptors, Peptide
dc.subjectRibonucleoproteins
dc.subjectSignal Recognition Particle
dc.subjectSuccinimides
dc.subjectViral Envelope Proteins
dc.subjectBiochemistry
dc.subjectCell Biology
dc.subjectMolecular Biology
dc.titleER translocation intermediates are adjacent to a nonglycosylated 34-kD integral membrane protein
dc.typeJournal Article
dc.source.journaltitleThe Journal of cell biology
dc.source.volume114
dc.source.issue1
dc.identifier.legacyfulltexthttps://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=1963&amp;context=oapubs&amp;unstamped=1
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/oapubs/964
dc.identifier.contextkey659145
refterms.dateFOA2022-08-23T16:55:03Z
html.description.abstract<p>We have used the homobifunctional cross-linking reagent disuccinimidyl suberate (DSS) to identify proteins that are adjacent to nascent polypeptides undergoing translocations across mammalian rough ER. Translocation intermediates were assembled by supplementing cell free translations of truncated mRNAs with the signal recognition particle (SRP) and microsomal membrane vesicles. Two prominent cross-linked products of 45 and 64 kD were detected. The 64-kD product was obtained when the cell free translation contained SRP, while formation of the 45-kD product required both SRP and translocation competent microsomal membrane vesicles. In agreement with previous investigators, we suggest that the 64-kD product arises by cross-linking of the nascent polypeptide to the 54-kD subunit of SRP. The 45-kD product resists alkaline extraction from the membrane, so we conclude that the 11-kD nascent polypeptide has been crosslinked to an integral membrane protein of approximately 34 kD (imp34). The cross-linked product does not bind to ConA Sepharose, nor is it sensitive to endoglycosidase H digestion; hence imp34 is not identical to the alpha or beta subunits of the signal sequence receptor (SSR). We propose that imp34 functions in concert with SSR to form a translocation site through which nascent polypeptides pass in traversing the membrane bilayer of the rough endoplasmic reticulum.</p>
dc.identifier.submissionpathoapubs/964
dc.contributor.departmentDepartment of Biochemistry and Molecular Biology
dc.source.pages21-33


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