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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
14
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pubmed:dateCreated |
1994-5-5
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pubmed:abstractText |
There are six intramolecular disulfide (S-S) bonds that form during intracellular folding of the human chorionic gonadotropin (hCG)-beta subunit. Site-directed mutagenesis of every pair of Cys residues involved in the formation of each S-S bond was used to examine the roles that S-S bonds play in beta subunit folding and secretion. Tryptic maps of secreted hCG-beta showed that only one S-S bond formed in all S-S bond mutants that failed to fold from the earliest detectable beta folding intermediate, p beta 1, into a second major intermediate, p beta 2 (C34A-C88A, C38A-C57A or C9A-C90A mutants), whereas all 5 remaining S-S bonds formed in mutants when p beta 1-->p beta 2 conversion occurred (C23A-C72A, C93A-C100A, or C26A-C110A mutants). Nonreducing SDS-polyacrylamide gel electrophoresis showed that beta multimers were secreted from cells expressing S-S bond mutations where the folding of p beta 1-->p beta 2 was blocked. However, for mutations where p beta 1-->p beta 2 conversion was efficient, beta monomers rather than multimers were secreted. For all cell lines studied, secreted hCG-beta migrated as monomeric beta during reducing SDS-polyacrylamide gel electrophoresis, indicating that hCG-beta multimers formed via intermolecular cross-linking of unpaired thiols. Tryptic maps of hCG-beta isolated from mutants lacking the 34-88 bond, where > 80% turnover occurs, showed that only the 38-57 S-S bond formed. beta Subunits lacking the 9-90 linkage also have only S-S bond 38-57 formed, but < 10% turnover of C9A-C90A hCG-beta occurs. Thus, subtle conformational differences between partially folded or misfolded beta subunits may determine whether hCG-beta is degraded, or undergoes intracellular translocation and secretion.
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pubmed:grant | |
pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Apr
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pubmed:issn |
0021-9258
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
8
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pubmed:volume |
269
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pubmed:geneSymbol |
hCG-&bgr;
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
10574-80
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pubmed:dateRevised |
2007-11-14
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pubmed:meshHeading |
pubmed-meshheading:8144645-Animals,
pubmed-meshheading:8144645-CHO Cells,
pubmed-meshheading:8144645-Chorionic Gonadotropin,
pubmed-meshheading:8144645-Chromatography, High Pressure Liquid,
pubmed-meshheading:8144645-Chromatography, Ion Exchange,
pubmed-meshheading:8144645-Cricetinae,
pubmed-meshheading:8144645-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:8144645-Humans,
pubmed-meshheading:8144645-Mutagenesis, Site-Directed,
pubmed-meshheading:8144645-Peptide Mapping,
pubmed-meshheading:8144645-Protein Folding,
pubmed-meshheading:8144645-Transfection,
pubmed-meshheading:8144645-Trypsin
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pubmed:year |
1994
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pubmed:articleTitle |
Misfolded human chorionic gonadotropin beta subunits are secreted from transfected Chinese hamster ovary cells.
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pubmed:affiliation |
Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha 68198.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
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