Characterization of a tachykinin peptide NK2 receptor transfected into murine fibroblast B82 cells.

Abstract

Membranes isolated from a murine fibroblast B82 cell line (SKLKB82#3) transfected with the bovine stomach cDNA pSKR56S exhibited binding of [His('25I) Considerable physiological and biochemical evidence exists for distinct NK1-, NK2-, and NK3-tachykinin peptide receptors. The preferred endogenous ligand appears to be substance P (SP) for NK1 receptors, neurokinin A (NKA) for NK2 receptors, and neurokinin B for NK3 receptors (1-4). Nakanishi and coworkers have cloned and expressed in Xenopus oocytes three tachykinin receptors-the rat brain NK1 receptor, the rat brain NK3 receptor, and the bovine stomach NK2 receptor (5-8). A functional cDNA encoding the rat SP receptor was also recently characterized by Hershey and Krause (9). The NK2 receptor has a calculated molecular mass of 43,066 Da, and all three receptors appear to be putative guanine nucleotide-binding protein (G protein)-linked receptors, each with seven hydrophobic, membranespanning domains, an extracellular amino terminus, and a cytoplasmic carboxyl terminus.The primary structures of numerous neurotransmitter receptors have been determined, but few of these are neuropeptide receptors (10,11). Recently, the bovine stomach cDNA, pSKR56S, was transfected into the murine fibroblast B82 cell line to produce cells (SKLKB82#3) with a high

Comparison of tachykinin NK1 and NK2 receptors in the circular muscle of the guinea‐pig ileum and proximal colon

“…For NK2-receptors, certain peptide (e.g. MEN 10,376, Maggi et al, 1991a) and nonpeptide antagonists (SR 48,968;Emonds-Alt et al, 1992; are more potent at human, bovine, guinea-pig and rabbit NK2 receptors than at rat or hamster NK2 receptors, while the cyclic peptide antagonist, L 659,877 (Williams et al, 1988) or the cyclic pseudopeptide MEN 10,573 (Quartara et al, 1992a) present the converse pattern of affinities (Maggi et al, 1990b;Van Giersbergen et al, 1991).…”

Tachykinin antagonists and capsaicin‐induced contraction of the rat isolated urinary bladder: evidence for tachykinin‐mediated cotransmission

“…The amino acid sequences of TK antagonists studied are shown in Table 1: spantide is a well known SP antagonist with some selectivity (about 10 times) for NK1 over NK2 receptors (Buck & Shatzer, 1988) Maggi et al, 1990a) in which Arg in position 11 has been replaced by Lys. Both MEN 10,207 and MEN 10,376 are highly selective NK2 receptor antagonists which discriminate between putative NK2 receptor subtypes (Maggi et al, 1990a;Van Giesbergen et al, 1991;Maggi et al, unpublished data). In preliminary experiments we found that MEN 10,207 antagonized the response to capsaicin of the rat isolated bladder but only at concentrations (10-30,UM) at which the peptide produced some agonist effect of its own.…”

Tachykinin receptors in the guinea‐pig renal pelvis: activation by exogenous and endogenous tachykinins

“…by GR 82,334, a selective NKI receptor antagonist (Hagan et al, 1991). In the past two years, pharmacological evidence has been provided for the heterogeneity of the tachykinin NK2 receptor 1991a;Buck et al, 1990;Van Giersbergen et al, 1991;Patacchini et al, 1991). Two criteria which allow the distinction between NK2 receptor subtypes have emerged: (a) rank order of potency of antagonists and (b) the different agonist/antagonists character of MDL 28,564.…”

“…However, the receptors involved in tachykininergic control of renal pelvis motility have not been fully characterized. Furthermore, recent studies have provided evidence for the heterogeneity of tachykinin NK2 receptors, possibly reflecting the existence of NK2 receptor subtypes 1991a;Buck et al, 1990;Van Giersbergen et al, 1991;Patacchini et al, 1991). The aim of this study was twofold: (a) to assess the type of tachykinin receptors mediating contraction of the guinea-pig isolated renal pelvis and (b) to assess the relative contribution of different tachykinin receptors to the response produced by endogenous tachykinins released during nerve stimulation.…”