Kassina senegalensis skin tachykinins: molecular cloning of kassinin and (Thr2, Ile9)-kassinin biosynthetic precursor cDNAs and comparative bioactivity of mature tachykinins on the smooth muscle of rat urinary bladder.
Tachykinins are among the most widely-studied families of regulatory peptides characterized by a highly-conserved C-terminal -Phe-X-Gly-Leu-Met.amide motif, which also constitutes the essential bioactive core. The amphibian skin has proved to be a rich source of these peptides with physalaemin from the skin of Physalaemus fuscomaculatus representing the archetypal aromatic tachykinin (X=Tyr or Phe) and kassinin from the skin of Kassina senegalensis representing the archetypal aliphatic tachykinin in which X=Val or Ile. Despite the primary structures of both mature peptides having been known for at least 30 years, neither the structures nor organizations of their biosynthetic precursors have been reported. Here we report the structure and organization of the biosynthetic precursor of kassinin deduced from cDNA cloned from a skin secretion library. In addition, a second precursor cDNA encoding the novel kassinin analog (Thr(2), Ile(9))-kassinin was identified as was the predicted mature peptide in skin secretion. Both transcripts exhibited a high degree of nucleotide sequence similarity and of open-reading frame translated amino acid sequences of putative precursor proteins. The translated preprotachykinins each consisted of 80 amino acid residues encoding single copies of either kassinin or its site-substituted analog. Synthetic replicates of each kassinin were found to be active on rat urinary bladder smooth muscle at nanomolar concentrations. The structural organization of both preprotachykinins differs from that previously reported for those of Odorrana grahami skin indicating a spectrum of diversity akin to that established for amphibian skin preprobradykinins.
Wang
Lei
L
Molecular Therapeutics Research, School of Pharmacy, Queen's University, Medical Biology Center, Belfast, Northern Ireland, UK.
Zhou
Mei
M
Lynch
Laura
L
Chen
Tianbao
T
Walker
Brian
B
Shaw
Chris
C
D000595:Amino Acid Sequence; D000818:Animals; D001001:Anura / Q000378:metabolism*; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D066298:In Vitro Techniques; D015286:Kassinin / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology*; D008297:Male; D008969:Molecular Sequence Data; D009130:Muscle, Smooth / Q000187:drug effects*; D051381:Rats; D016415:Sequence Alignment; D012867:Skin / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D015320:Tachykinins / Q000138:chemical synthesis / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D001743:Urinary Bladder / Q000187:drug effects*
D018076:DNA, Complementary; D015320:Tachykinins; D015286:Kassinin