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Primary structure of fox ( Vulpes vulpes ) proinsulin based on sequence studies of pancreatic peptides and cDNA
Author(s) -
Fiertek Dariusz,
Gromowska Małgorzata,
Andersen Asser S.,
Hansen Per H.,
Majewski Tadeusz,
Izdebski Jan
Publication year - 2000
Publication title -
journal of peptide science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 66
eISSN - 1099-1387
pISSN - 1075-2617
DOI - 10.1002/1099-1387(200008)6:8<413::aid-psc268>3.0.co;2-z
Subject(s) - vulpes , proinsulin , complementary dna , protein primary structure , sequence (biology) , peptide sequence , computational biology , biology , microbiology and biotechnology , chemistry , biochemistry , endocrinology , gene , ecology , insulin , predation
Insulin and C‐peptide were extracted and purified from fox ( Vulpes vulpes ) pancreas using gel filtration, ion‐exchange chromatography and HPLC. Chromatographic data for the insulin, as well as for its oxidized and carboxymethylated chains proved it to be identical to that of polar fox ( Alopex lagopus ) and dog. The sequence analysis of a peptide which was assumed to be the corresponding C‐peptide revealed that it comprises 23 amino acid residues and is identical to the C‐peptide fragment isolated from dog pancreas; it differs from polar fox C‐peptide by a single substitution (Asp→Glu). mRNA was isolated from pancreatic tissue and cDNA was obtained by reverse transcription. A polymerase chain reaction was performed using gene‐specific primers to obtain a DNA fragment corresponding to part of fox proinsulin. DNA sequencing revealed 100% identity to dog proinsulin at the protein level, although some amino acids were encoded by different codons. The total sequence of proinsulin was deduced from these results. Copyright © 2000 European Peptide Society and John Wiley & Sons, Ltd.