Anti-bovine rhodopsin monoclonal antibody recognizing light-dependent structural change.
The antigenic structure of the bovine rhodopsin molecule was investigated by using a bovine rhodopsin-specific monoclonal antibody designated Rh 29. Competition assay with sealed intact disks and broken disks indicated that the antibody-binding region was localized in the intradiscal surface. An antigenic peptide obtained by a cyanogene bromide cleavage of rhodopsin was purified and determined as residues 2-39 in the amino acid sequence. Further analysis suggested that the antigenic determinant included at least residues 21-25. These results were consistent with the structural model for membrane topology of rhodopsin. The antigenicity of the rhodopsin was compared among several states. The antibody bound to both ammonyx LO-solubilized unbleached and bleached rhodopsin. In contrast, upon membrane-embedded rhodopsin, unbleached one was 100-times less antigenic than bleached one. The results suggested that the segment around the determinant of membrane-embedded rhodopsin should undergo a structural change upon absorption of light. Rh 29 detected a band corresponding to bovine, porcine and octopus opsins in immunoblotting. Protein blot of crayfish rhabdome did not show any reactive band. These bands except for crayfish reacted with concanavalin A as well. The N-terminal structure may, therefore, conserved between mammal and erthropoda and diverge between them and cepharopoda.
Takao
Masashi
M
Department of Molecular Genetics, Institute of Development, Aging and Cancer, Tohoku University, Sendai, Miyagi 980-8575, Japan.
Iwasa
Tatsuo
T
Yamamoto
Hiroaki
H
Takeuchi
Takuji
T
Tokunaga
Fumio
F
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D003400:Astacoidea; D001666:Binding Sites, Antibody; D002417:Cattle; D003208:Concanavalin A / Q000276:immunology; D017124:Conserved Sequence; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D015151:Immunoblotting; D007700:Kinetics; D008027:Light; D008969:Molecular Sequence Data; D049831:Octopodiformes; D011487:Protein Conformation / Q000528:radiation effects*; D012243:Rhodopsin / Q000737:chemistry* / Q000276:immunology*; D012995:Solubility; D013552:Swine
D000911:Antibodies, Monoclonal; D000939:Epitopes; D003208:Concanavalin A; D012243:Rhodopsin