Characterization of a tachykinin peptide NK sub 2 receptor transfected into murine fibroblast B82 cells

Journal Article · Fri Mar 01 04:00:00 EST 1991 · Proceedings of the National Academy of Sciences of the United States of America; (United States)

DOI:https://doi.org/10.1073/pnas.88.5.1661· OSTI ID:6203361

Van Giersbergen, P L.M. ^[1]; Shatzer, S A; Buck, S H ^[2]; Henderson, A K; Lai, J; Nakanishi, Shigetada ^[3]

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1. Marion Merrell Dow Research Inst., Cincinnati, OH (United States) Univ. of Cincinnati, OH (United States)
2. Marion Merrell Dow Research Inst., Cincinnati, OH (United States)
3. Kyoto Univ. (Japan)

Membranes isolated from a murine fibroblast B82 cell line (SKLKB82{number sign}3) transfected with the bovine stomach cDNA pSKR56S exhibited binding of (His({sup 125}I){sup 1})neurokinin A ({sup 125}I-NKA) to a single population of sites with a B{sub max} of 147 fmol/mg of protein and a K{sub d} of 0.59 nM. The ligand binding in SKLKB82{number sign}3 cells was reversible. Thus, SKLKB82{number sign}3 cells have been transfected with NK{sub 2} receptors that have become associated with an endogenous guanine nucleotide-binding protein. In comparison with membranes from the hamster urinary bladder, a tissue enriched in NK{sub 2} receptors, NK{sub 2} receptor antagonists displayed markedly different potencies, either more or less potent, in inhibiting specific binding in membranes of the transfected cells. Furthermore, inhibition of {sup 125}I-NKA binding by nucleotide analogues was markedly different in SKLKB82{number sign}3 cells compared with hamster bladder tissue. The different binding profile in the cells is not due to an artefact introduced during cDNA transfection because a similar profile was also observed in bovine stomach membranes. These results may indicate the existence of two distinct NK{sub 2} receptors.

• OSTI ID: 6203361
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (United States), Journal Name: Proceedings of the National Academy of Sciences of the United States of America; (United States) Vol. 88:5; ISSN 0027-8424; ISSN PNASA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMAL CELLS
ANIMALS
AUTONOMIC NERVOUS SYSTEM AGENTS
BETA DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
CATIONS
CELL CONSTITUENTS
CELL MEMBRANES
CHARGED PARTICLES
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
DNA
DRUGS
ELECTRON CAPTURE RADIOISOTOPES
FIBROBLASTS
HAMSTERS
INTERMEDIATE MASS NUCLEI
INTERNAL CONVERSION RADIOISOTOPES
IODINE 125
IODINE ISOTOPES
IONS
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
KININS
MAMMALS
MEMBRANE PROTEINS
MEMBRANES
MICE
NEUROREGULATORS
NUCLEI
NUCLEIC ACIDS
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
PEPTIDES
POLYPEPTIDES
PROTEINS
RADIOISOTOPES
RADIORECEPTOR ASSAY
REACTION KINETICS
RECEPTORS
RECOMBINANT DNA
RODENTS
SOMATIC CELLS
TRACER TECHNIQUES
VERTEBRATES