Structure of a biologically active neurotensin-related peptide obtained from pepsin-treated albumin(s).
Using a radioimmunoassay toward the COOH-terminal region of neurotensin, an immunoreactive and biologically active neurotensin-related peptide (NRP) has been isolated from pepsin-treated fractions of bovine, canine, human, and rat plasma. Bovine NRP was identified as H-Ile-Ala-Arg-Arg-His-Pro-Tyr-Phe-Leu-OH, which is similar in structure to both neurotensin and angiotensin I. Canine and human NRP also had the above amino acid composition, whereas that obtained from rat plasma had valine substituted for isoleucine. At their concentrations in pepsin-treated plasmas (2-6 microM) rat, human and canine NRP were shown to increase vascular permeability when injected intradermally into rats and to release histamine from rat mast cells in vitro. The pure peptides also cross-reacted very effectively at nanomolar concentrations in a radioreceptor assay for neurotensin. The protein(s) which liberated NRP upon pepsin treatment were purified about 7-fold and shown to behave like albumin during sodium dodecyl sulfate-polyacrylamide gel electrophoresis, isoelectric focusing, and high pressure liquid chromatography on muBondapak C4. In addition, the purified preparations were found to react with anti-albumin antisera during immunodiffusion. Although the amino acid sequence of NRP was not found in albumin, a partial sequence homology was noted for NRP and various segments of bovine albumin. Using V8 protease, glutamyl residues were shown to lie within 3-4 amino acids of each end of NRP, as also occurs for the related segments in albumin. These results suggest that a subset of albumin-related protein(s) could serve as precursor(s) to biologically active neurotensin-related peptide(s).
D000595:Amino Acid Sequence; D000818:Animals; D001681:Biological Assay; D001920:Bradykinin / Q000494:pharmacology; D002417:Cattle; D004285:Dogs; D004305:Dose-Response Relationship, Drug; D010450:Endopeptidases / Q000378:metabolism; D006636:Histamine Release / Q000187:drug effects; D006801:Humans; D008407:Mast Cells / Q000187:drug effects; D009496:Neurotensin / Q000032:analysis*; D010434:Pepsin A / Q000494:pharmacology*; D051381:Rats; D012697:Serine Endopeptidases; D012709:Serum Albumin / Q000032:analysis*
D012709:Serum Albumin; D009496:Neurotensin; D010450:Endopeptidases; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase; D010434:Pepsin A; D001920:Bradykinin