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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
1
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pubmed:dateCreated |
1976-10-29
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pubmed:abstractText |
A novel manual method for protein-sequence analysis is described. Three peptides, the hexapeptide (Leu-TRP-Met-Arg-Phe-Ala), insulin A chain and glucagon were used to test this technique. Peptides (1 or 2 nmol) were hydrolysed with acid and their qualitative amino acid compositions were confirmed by reacting with 4-NN-dimethylaminoazobenzene-4'-sulphonylchloride and 4-NN-dimethylaminoazobenzene 4'-isothiocyanate. Sequence determination of 20-200 nmol of peptide was then performed by the combined use of phenyl isothiocyanate and 4-NN-dimethylaminoazobenzene 4'-isothiocyanate, a new procedure that is analogous to the dansyl-Edman method with the replacement of dansyl chloride by 4-NN-dimethylaminoazobenzene 4'-isothiocyanate as the N-terminal residue determination reagent. On t.l.c. this new N-terminal reagent gave brightly coloured 4-NN-dimethylaminoazobenzene-4-thiohydantoins of amino acids and showed the following advantages: (1) the detection sensitivity is in the pmol range; (2) u.v. observation is not required; (3) there is no destruction of acid-labile amino acids; (4) two-dimensional t.l.c. separation is adequate to identify 24 amino acids, except leucine and isoleucine (this pair of amino acids can be resolved by using 4-NN-dimethylaminoazobenzene-4'-sulphonyl chloride); (5) the determination of a new N-terminal residue (from coupling to t.l.c. identification) takes only 3 h; (6) the colour difference beteen isothiocyanate, thiocarbamoyl and thiohydantoin derivatives facilitates the identifications.
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/822842-13831429,
http://linkedlifedata.com/resource/pubmed/commentcorrection/822842-4716391,
http://linkedlifedata.com/resource/pubmed/commentcorrection/822842-4838784,
http://linkedlifedata.com/resource/pubmed/commentcorrection/822842-4923920,
http://linkedlifedata.com/resource/pubmed/commentcorrection/822842-5061852,
http://linkedlifedata.com/resource/pubmed/commentcorrection/822842-5578618,
http://linkedlifedata.com/resource/pubmed/commentcorrection/822842-6059350,
http://linkedlifedata.com/resource/pubmed/commentcorrection/822842-808149,
http://linkedlifedata.com/resource/pubmed/commentcorrection/822842-812880
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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/Glucagon,
http://linkedlifedata.com/resource/pubmed/chemical/Insulin,
http://linkedlifedata.com/resource/pubmed/chemical/Peptides,
http://linkedlifedata.com/resource/pubmed/chemical/Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/p-Dimethylaminoazobenzene
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pubmed:status |
MEDLINE
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pubmed:month |
Jul
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pubmed:issn |
0264-6021
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
1
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pubmed:volume |
157
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
77-85
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pubmed:dateRevised |
2011-11-17
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pubmed:meshHeading |
pubmed-meshheading:822842-Amino Acid Sequence,
pubmed-meshheading:822842-Chromatography, Thin Layer,
pubmed-meshheading:822842-Glucagon,
pubmed-meshheading:822842-Insulin,
pubmed-meshheading:822842-Methods,
pubmed-meshheading:822842-Peptides,
pubmed-meshheading:822842-Proteins,
pubmed-meshheading:822842-p-Dimethylaminoazobenzene
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pubmed:year |
1976
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pubmed:articleTitle |
A novel manual method for protein-sequence analysis.
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pubmed:publicationType |
Journal Article
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