Proenkephalin A processing in the upper digestive tract: isolation and characterisation of phosphorylated N-terminally extended Met-enkephalin Arg6Phe7 variants.
Previous studies suggest the processing of proenkephalin A in the porcine upper digestive tract might differ from that in the brain. To characterise more precisely some of the products, we have used antibodies to Met-enkephalin Arg6Phe7 (MERF) in radioimmunoassay to monitor the isolation of immunoreactive peptides from extracts of porcine pyloric antral muscle, antral mucosa, and duodenum. Sephadex G50 gel filtration of each extract produced a single broad peak of high-molecular-weight MERF-immunoreactivity. On anion-exchange chromatography the antral muscle MERF-immunoreactivity fractionated into two major peaks, and that from the antral mucosa and duodenum each into four major peaks, suggesting tissue specific processing of proenkephalin A within the porcine gut. Reverse-phase HPLC and Edman degradation analysis revealed that the least acidic antral muscle peptide was a 31-residue N-terminally extended form of MERF that is equivalent to proenkephalin A 209-239. Alkaline phosphatase digestion of the N-terminally extended MERF variants indicated that some of these peptides were modified by phosphorylation. We conclude that there are complex patterns of proenkephalin A processing in the porcine gut, which in part are due to phosphorylation.
MRC Secretory Control Research Group, University of Liverpool, England, U.K.
A
A
Watkinson
G J
GJ
Dockray
J
J
Young
H
H
Gregory
D000469:Alkaline Phosphatase / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004386:Duodenum / Q000378:metabolism*; D004744:Enkephalin, Methionine / Q000031:analogs & derivatives* / Q000302:isolation & purification / Q000378:metabolism; D004745:Enkephalins / Q000378:metabolism*; D005753:Gastric Mucosa / Q000378:metabolism*; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011498:Protein Precursors / Q000378:metabolism*; D011706:Pyloric Antrum / Q000378:metabolism; D011863:Radioimmunoassay; D013552:Swine
D004745:Enkephalins; D011498:Protein Precursors; C029992:proenkephalin; D004744:Enkephalin, Methionine; C027511:enkephalin-Met, Arg(6)-Phe(7)-; D000469:Alkaline Phosphatase