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Pancreatic polypeptide from rat pancreas.

J R Kimmel, H G Pollock, R E Chance

    Endocrinology
    |May 1, 1984
    PubMed
    Summary

    Researchers isolated and characterized rat pancreatic polypeptide (PP), a 36-amino acid peptide. They determined its amino acid sequence and identified oxidation of a methionine residue, which affects its immunoactivity.

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    Area of Science:

    • Endocrinology and Metabolism
    • Protein Chemistry
    • Bioanalytical Chemistry

    Background:

    • Pancreatic polypeptide (PP) is a peptide hormone with diverse physiological roles.
    • Understanding the structure and properties of PP from different species is crucial for comparative studies.
    • Previous research has characterized PP from various mammalian sources, but rat PP required detailed investigation.

    Purpose of the Study:

    • To isolate and purify pancreatic polypeptide (PP) from rat pancreas.
    • To determine the amino acid composition and sequence of rat PP.
    • To investigate the stability and immunoactivity of rat PP, particularly concerning methionine oxidation.

    Main Methods:

    • Isolation of rat PP using sequential chromatography: gel filtration, ion exchange, and high-performance liquid chromatography (HPLC).

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  • Radioimmunoassay (RIA) utilizing an antibody against the carboxyl-terminal hexapeptide of bovine PP for monitoring isolation.
  • Amino acid analysis and sequencing to determine the primary structure of rat PP.
  • Chemical reduction with mercaptoethanol to reverse methionine oxidation.

    • Main Results:

    • Rat PP was successfully isolated and purified, yielding approximately 2 mg/kg of pancreatic tissue.
    • Rat PP consists of 36 amino acids with a composition similar to other mammalian PPs.
    • The identified amino acid sequence is Ala-Pro-Leu-Glu-Pro-Met-Tyr-Pro-Gly-Asp-Tyr-Ala-Thr-His-Glu-Gln-Arg-Ala-Gln-Tyr-Glu-Thr-Gln-Leu-Arg-Arg-Tyr-Ile-Asn-Thr-Leu-Thr-Arg-Pro-Arg-Tyr-NH2.
    • A single methionine residue is prone to oxidation, forming two immunoactive peaks in HPLC, which can be reversed by reduction.

    Conclusions:

    • The study provides the complete amino acid sequence of rat pancreatic polypeptide.
    • The determined sequence maintains structural features essential for the globular conformation of PP.
    • Methionine oxidation is a significant factor affecting rat PP's stability and detection, requiring consideration in experimental procedures.