Isolation and quantitation of several new peptides from the canine neurotensin/neuromedin N precursor.
Using antisera towards the bioactive peptides, neurotensin and neuromedin N, as well as towards the N-terminal and C-terminal regions of their shared 170-residue precursor, peptides representing various portions of the precursor were isolated from extracts of canine ileum. In total, seven peptides were isolated, two of which had not been previously identified. One was the C-terminal tail of the precursor (Gly-Ser-Tyr-Tyr-Tyr) and the other was the tail peptide extended N-terminally to include neurotensin (Glp-Leu-Tyr-Glu-Asn-Lys-Pro-Arg-Arg-Pro-Tyr-Ile-Leu-Lys-Arg-Gly-Ser-Tyr -Tyr- Tyr). By comparing the measured concentrations for each of the identified peptides, it was established that processing at the three Lys-Arg cleavage sites within the precursor did not occur to the same extent. Cleavage at the N-terminus of neuromedin N was approximately 10% complete, that between neurotensin and the tail was approximately 90% complete, and that between neuromedin N and neurotensin was approximately 95% complete. Three immunoreactive proteins were also identified by immunochemical and chromatographic analyses: N-terminally extended neuromedin N (125 residues), N-terminally extended neurotensin (140 residues), and the entire 147-residue precursor. It was concluded that neurotensin, tail and large molecular neuromedin N were the primary products of processing for this precursor in canine ileum, while minor products included neuromedin N, neurotensin tail, and large molecular neurotensin.
Carraway
R E
RE
Department of Physiology, University of Massachusetts Medical School, Worcester 01655.
Mitra
S P
SP
Salmonsen
R
R
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004285:Dogs; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009496:Neurotensin / Q000737:chemistry*; D010446:Peptide Fragments / Q000737:chemistry* / Q000302:isolation & purification*; D011498:Protein Precursors / Q000737:chemistry*; D011863:Radioimmunoassay
D000596:Amino Acids; D010446:Peptide Fragments; D011498:Protein Precursors; C042090:neuromedin N; D009496:Neurotensin