The complete primary structure of pancreatic polypeptide from the European common frog, Rana temporaria.
Using an antiserum directed against the highly-conserved C-terminal hexapeptide amide of mammalian pancreatic polypeptide (PP), numerous immunoreactive endocrine cells were identified within the pancreas of the European common frog, R. temporaria. An acidified ethanolic extract of pancreatic tissue (0.859 g, n = 35) contained 26.2 nmol equivalents/g of tissue. Gel permeation chromatography of the extract resolved a single peak of immunoreactivity co-eluting with synthetic bovine PP standard. Reverse phase HPLC of this material resolved a single peak of immunoreactivity which was purified to homogeneity following chromatography on a semipreparative C-18 column and an analytical C-8 column. Plasma desorption mass spectrometry (PDMS) of the purified peptide resolved a single component with a molecular mass of 4240.9 Da. Direct gas phase sequencing established the sequence of the first 26 residues. Following incubation of the peptide with endopeptidase Asp-N and direct application of the digest to the sequencer, the entire primary structure of the peptide was established as: APSEPHHPGDQATQDQLAQYYSDLYQYITFVTRPRF. The derived molecular mass of this peptide, incorporating a C-terminal amide, was 4240.6 Da which is entirely consistent with that obtained by PDMS.
McKay
D M
DM
Department of Medicine, Queen's University of Belfast, Northern Ireland.
Shaw
C
C
Thim
L
L
Johnston
C F
CF
Halton
D W
DW
Fairweather
I
I
Buchanan
K D
KD
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D010191:Pancreatic Polypeptide / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification; D011896:Rana temporaria
D000596:Amino Acids; D010191:Pancreatic Polypeptide