| Identification of the C-terminally alpha-amidated amino acid in peptides by high-performance liquid chromatography. | |
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MedLine Citation:
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PMID: 3549288 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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A sensitive method for the rapid identification of the C-terminally amidated amino acid in peptides is described. Peptides containing the alpha-amide group at the C-terminus were cleaved with endopeptidases. The fragments released (oligopeptides, amino acids and the C-terminally amidated residue) are coupled to phenylisothiocyanate. The phenylthiocarbamoyl derivative of the amino acid alpha-amide is selectively extracted from the mixture by alkaline butyl acetate and identified by a high-performance liquid chromatography system that enables rapid and complete separation of the derivatives of 17 amino acid amides at a detection limit of 20-50 pmol. The C-terminal alpha-amides of neurokinin-A (Met-NH2), mammalian secretin (Val-NH2), pancreatic polypeptide (Tyr-NH2) and peptide HI (Ile-NH2) are unequivocally determined at a level of 0.5-2 nmol per peptide. This method was used to characterize a crude peptide fraction prepared from porcine brain. Cholecystokinin-58 was identified in this fraction by detection of phenylthiocarbamoyl-phenylalaninamide. The method is suitable for the identification of the C-terminal alpha-amidated residue of purified peptides, but can also be used as a screening strategy to isolate from complex biological extracts novel peptides containing an alpha-amidated amino acid at the C-terminus. |
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Authors:
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W E Schmidt; J M Conlon; V Mutt; M Carlquist; B Gallwitz; W Creutzfeldt |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: European journal of biochemistry / FEBS Volume: 162 ISSN: 0014-2956 ISO Abbreviation: Eur. J. Biochem. Publication Date: 1987 Feb |
Date Detail:
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Created Date: 1987-05-01 Completed Date: 1987-05-01 Revised Date: 2007-07-23 |
Medline Journal Info:
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Nlm Unique ID: 0107600 Medline TA: Eur J Biochem Country: GERMANY, WEST |
Other Details:
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Languages: eng Pagination: 467-72 Citation Subset: IM |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Amides
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analysis* Amino Acids / analysis* Animals Brain Chemistry Chromatography, High Pressure Liquid Digestive System / analysis Endopeptidases Hydrolysis Peptides / analysis* Swine |
| Chemical | |
Reg. No./Substance:
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0/Amides; 0/Amino Acids; 0/Peptides; EC 3.4.-/Endopeptidases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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