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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

Evidence of NK1 and NK2 tachykinin receptors and their involvement in histamine release in a murine mast cell line.

Binding of [3H]substance P (SP) and histamine release were examined using a cloned mouse mast cell line. SP binding was saturable and specific. In the presence of 30 mM Na2SO4/50 mM Tris buffer, SP interacted with two types of binding sites with Kd values of 0.3 and 40 nM. High-affinity SP binding was blocked by the inclusion of 0.5 uM of the NK1 receptor selective ligand septide in the binding mixture. Neurokinin A ( NKA) evoked concentration-dependent histamine release. At concentrations in the nanomolar range, the NK1 preferring agonists SP, SP methylester and physalaemin evoked less than or equal to 5% net release of histamine, which was substantially less than the maximum effect of NKA (+37%) in the micromolar range. Pretreatment of the cells with the NK2 antagonist peptide A reduced NKA-induced histamine release. [D-Arg1,D-Phe5,D-Trp7,9,Leu11]-substance P, a putative SP antagonist, also elicited histamine release in the micromolar range, apparently acting as an agonist at the NK2 site. Compound 48/80, N-terminal SP fragments, neurokinin B and the two selective NK2 receptor antagonists cyclo(Gln-Trp-Phe-(R)-[ANC-2]Leu-Met) (peptide A) and cyclo(Gln-Trp-Phe-Gly-Leu-Met) (peptide B) were ineffective. Although the results suggest the coexistence of functional NK1 and NK2 receptors, it appears that in this mast cell line neurokinin- induced histamine release is primarily mediated by the NK2 receptor, characterized biochemically as a low affinity binding site with a Kd value of 40 nM for SP.[1]


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