Purification and characterization of insulin and the C-peptide of proinsulin from Przewalski's horse, zebra, rhino, and tapir (Perissodactyla).
Within the order Perissodactyla, the primary structure of insulin has been strongly conserved. Insulin from Przewalski's horse and the mountain zebra (suborder Hippomorpha) is the same as that from the domestic horse and differs from insulin from the white rhinoceros and mountain tapir (suborder Ceratomorpha) by a single substitution (Gly-->Ser) at position 9 in the A-chain. A second molecular form of Przewalski's horse insulin isolated in this study was shown to represent the gamma-ethyl ester of the Glu17 residue of the A-chain. This component was probably formed during the extraction of the pancreas with acidified ethanol. The amino acid sequence of the C-peptide of proinsulin has been less well conserved. Zebra C-peptide comprises 31 amino acid residues and differs from Przewalski's horse and domestic horse C-peptide by one substitution (Gln30-->Pro). Rhino C-peptide was isolated only in a truncated form corresponding to residues (1-23) of intact C-peptide. Its amino acid sequence contains three substitutions compared with the corresponding region of horse C-peptide. It is postulated that the substitution (Pro23-->Thr) renders rhino C-peptide more liable to proteolytic cleavage by a chymotrypsin-like enzyme than horse C-peptide. C-peptide could not be identified in the extract of tapir pancreas, suggesting that proteolytic degradation may have been more extensive than in the rhino. In contrast to the ox and pig (order Artiodactyla), there was no evidence for the expression of more than one proinsulin gene in the species of Perissodactyla examined.
Henry
J S
JS
Department of Biomedical Sciences, Creighton University School of Medicine, Omaha, Nebraska 68178.
Lance
V A
VA
Conlon
J M
JM
D000595:Amino Acid Sequence; D000818:Animals; D002096:C-Peptide / Q000032:analysis / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid; D005260:Female; D006736:Horses; D007328:Insulin / Q000032:analysis / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D010527:Perissodactyla / Q000097:blood*; D011384:Proinsulin / Q000032:analysis / Q000302:isolation & purification*; D011863:Radioimmunoassay; D013045:Species Specificity
D002096:C-Peptide; D007328:Insulin; D011384:Proinsulin