A monoclonal antibody to the ligand-binding domain of the neurokinin 1 receptor (NK1-R) for the neuropeptide substance P.
Monoclonal antibodies to the binding site of the NK1 receptor for the neuropeptide substance P were produced in mice using the complementary or antisense peptide methodology. Among several anti-peptide monoclonal antibodies, we selected the mAb12 antibody which specifically crossreacted, through its paratope, with a binding site present on membranes from rat parotid gland cells, with an affinity close to 2 x 10(-7) M and with membranes from CHO cells expressing human brain NK1 receptors. Immunocytochemical investigations using mAb12 revealed immunostaining whose distribution in the dorsal horns of rat spinal cord fits well with the known location of NK1 receptors. In both biochemical and immunocytochemical experiments, the competition occurring between the antibody and substance P, or a substance P-protein conjugate, indicates that mAb12 recognizes a membrane epitope located at or near the substance P binding domain on the NK1 receptor. Immunization of mice with mAb12 led to the production of specific anti-substance P antibodies, again suggesting that mAb12 shares common structural features with the neuropeptide. This monoclonal antibody can now be used in further biochemical or cytochemical characterizations of NK1 receptors. Owing to its fine specificity, mAb12 could also serve as a molecular model for designing peptides, possibly displaying pharmacological properties in the various processes in which substance P is involved, e.g. immunomodulation, inflammation or chronic pain.
Déry
O
O
CEA, Service de Pharmacologie et d'Immunologie, DSV/DRM, Gif-sur-Yvette, France.
Frobert
Y
Y
Zerari
F
F
Créminon
C
C
Grassi
J
J
Fischer
J
J
Conrath
M
M
Couraud
J Y
JY
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001665:Binding Sites; D016466:CHO Cells; D006224:Cricetinae; D005260:Female; D006801:Humans; D007114:Immunization; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D051381:Rats; D017208:Rats, Wistar; D018040:Receptors, Neurokinin-1 / Q000032:analysis* / Q000276:immunology; D013373:Substance P / Q000276:immunology*
D000911:Antibodies, Monoclonal; D018040:Receptors, Neurokinin-1; D013373:Substance P