Primary structure and receptor-binding properties of a neurokinin A-related peptide from frog gut

Biochem J. 1992 Nov 1;287 ( Pt 3)(Pt 3):827-32. doi: 10.1042/bj2870827.

Abstract

A tachykinin peptide was isolated from an extract of the intestine of the European green frog, Rana ridibunda, and its primary structure was established as: His-Lys-Leu-Asp-Ser-Phe-Ile-Gly-Leu-Met.CONH2. This sequence was confirmed by chemical synthesis and shows two amino acid substitutions (leucine for threonine at position 3 and isoleucine for valine at position 7) compared with neurokinin A. Binding parameters for synthetic [Leu3,Ile7]neurokinin A and mammalian tachykinins were compared using receptor-selective radioligands and crude membranes from tissues enriched in the NK1, NK2 and NK3 receptors. [Leu3,Ile7]Neurokinin A was approx. 3-fold less potent than substance P in inhibiting the binding of 125I-labelled [Sar9,Met(O2)11]substance P (labelled with Bolton-Hunter reagent) to rat submandibular gland (NK1 receptor), 8-fold less potent than neurokinin A in inhibiting the binding of [2-[125I]iodohistidine1]neurokinin A to rat stomach fundus (NK2 receptor) and 6-fold less potent than neurokinin B in inhibiting the binding of 125I-Bolton-Hunter-labelled scyliorhinin II to rat brain (NK3 receptor). Thus the frog neurokinin A-related peptide shows moderate affinity but lack of selectivity for all three tachykinin-binding sites in rat tissues. This non-selectivity is similar to that displayed by the molluscan tachykinin, eledoisin, which also contains an isoleucine residue in the corresponding position in the molecule.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Binding, Competitive
  • Chromatography, Gel
  • Chromatography, High Pressure Liquid
  • Female
  • Intestine, Small / chemistry*
  • Molecular Sequence Data
  • Neurokinin A / analogs & derivatives*
  • Neurokinin A / chemistry
  • Neurokinin A / isolation & purification
  • Neurokinin A / metabolism
  • Rana ridibunda
  • Rats
  • Receptors, Neurokinin-2
  • Receptors, Neurotransmitter / metabolism*
  • Sequence Homology, Amino Acid
  • Tachykinins / metabolism

Substances

  • Receptors, Neurokinin-2
  • Receptors, Neurotransmitter
  • Tachykinins
  • neurokinin A, Leu(3)-Ile(7)-
  • Neurokinin A