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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
40
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pubmed:dateCreated |
1998-11-4
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pubmed:abstractText |
Purified rabbit and sheep sex hormone-binding globulins (SHBGs) were photolabeled by Delta 6-testosterone. The maximal levels of specific incorporation were respectively 0.33 and 0.30 mol of label/mol of homodimer. Tryptic cleavage of photolabeled SHBGs gave a single radioactive peptide for rabbit SHBG and two major radioactive peptides S1 and S2 for sheep SHBG. Edman sequencing of the photolabeled peptide of rabbit SHBG revealed a single sequence corresponding to peptidic fragment Leu-118-Lys-134. Subcleavage of this peptide with elastase led to a single radioactive peptidic fragment corresponding to dipeptide Met-133-Lys-134, identified by mass spectrometry, while deletion of the C-terminal residue with carboxypeptidase B showed that all the radioactivity remained on peptide Leu-118-Met-133, thus demonstrating that photolabeling occurred exclusively on Met-133, the only residue common to the two radioactive subcleaved peptides. Edman sequencing of peptides S1 and S2 of sheep SHBG showed a same single sequence corresponding to residues Gln-126-Arg-140 which contained no identifiable phenylthiohydantoin derivative at cycle 14, thus indicating that in both cases the corresponding Met-139 residue is the main site of photolabeling, as confirmed for peptide S1 by the presence at this cycle of a major peak of radioactivity while in peptide S2 the photoattachment of Delta 6-testosterone was found labile in the conditions of sequencing. The photolabeled peptide S1 was characterized by mass spectrometry which showed the covalent fixation of one mole of Delta 6-testosterone and the presence of a biantennary oligosaccharide attached at Asn-133, which suggests that the steroid-binding site is probably not deeply buried in the SHBG homodimer.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/6-dehydrotestosterone,
http://linkedlifedata.com/resource/pubmed/chemical/Methionine,
http://linkedlifedata.com/resource/pubmed/chemical/Peptide Fragments,
http://linkedlifedata.com/resource/pubmed/chemical/Photoaffinity Labels,
http://linkedlifedata.com/resource/pubmed/chemical/Sex Hormone-Binding Globulin,
http://linkedlifedata.com/resource/pubmed/chemical/Testosterone,
http://linkedlifedata.com/resource/pubmed/chemical/Trypsin
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pubmed:status |
MEDLINE
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pubmed:month |
Oct
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pubmed:issn |
0006-2960
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
6
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pubmed:volume |
37
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
14088-97
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:9760244-Amino Acid Sequence,
pubmed-meshheading:9760244-Animals,
pubmed-meshheading:9760244-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:9760244-Gamma Rays,
pubmed-meshheading:9760244-Hydrolysis,
pubmed-meshheading:9760244-Kinetics,
pubmed-meshheading:9760244-Methionine,
pubmed-meshheading:9760244-Molecular Sequence Data,
pubmed-meshheading:9760244-Peptide Fragments,
pubmed-meshheading:9760244-Photoaffinity Labels,
pubmed-meshheading:9760244-Rabbits,
pubmed-meshheading:9760244-Sequence Homology, Amino Acid,
pubmed-meshheading:9760244-Sex Hormone-Binding Globulin,
pubmed-meshheading:9760244-Sheep,
pubmed-meshheading:9760244-Testosterone,
pubmed-meshheading:9760244-Trypsin
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pubmed:year |
1998
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pubmed:articleTitle |
Photoaffinity labeling of homologous Met-133 and Met-139 amino acids of rabbit and sheep sex hormone-binding globulins with the unsubstituted Delta 6-testosterone photoreagent.
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pubmed:affiliation |
Institut National de la Santé et de la Recherche Médicale, Unité INSERM U 329, Pathologie Hormonale Moléculaire, Hôpital Debrousse, Lyon, France.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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