Characterization of the substance P (NK-1) receptor in tunicamycin-treated transfected cells using a photoaffinity analogue of substance P.
Chinese hamster ovary cells expressing the N-glycosylated substance P (NK-1) receptor were treated with the glycosylation inhibitor tunicamycin and photolabeled with 125I-Bolton-Hunter-p-benzoyl-L-phenylalanine8-substance P. Two radioactive proteins of M(r) 80,000 and 46,000, representing the glycosylated and nonglycosylated substance P (NK-1) receptor, respectively, were observed. The IC50 for the inhibition of photolabeling of both receptor forms was 0.3 +/- 0.1 nM for substance P and 30 +/- 5 nM for neurokinin A (substance K). Thus, glycosylation of the substance P (NK-1) receptor has no detectable effect on the affinity of the substance P (NK-1) receptor for substance P or neurokinin A (substance K).
Kage
R
R
Department of Pharmacology and Experimental Therapeutics, Boston University School of Medicine, MA 02118.
Hershey
A D
AD
Krause
J E
JE
Boyd
N D
ND
Leeman
S E
SE
D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel; D006031:Glycosylation; D008027:Light; D015288:Neurokinin A; D009844:Oligosaccharides / Q000032:analysis; D018040:Receptors, Neurokinin-1 / Q000032:analysis* / Q000378:metabolism*; D013373:Substance P / Q000031:analogs & derivatives* / Q000378:metabolism*; D014162:Transfection; D014415:Tunicamycin / Q000494:pharmacology*
D009844:Oligosaccharides; D018040:Receptors, Neurokinin-1; D014415:Tunicamycin; D013373:Substance P; D015288:Neurokinin A