Anti-Bovine Rhodopsin Monoclonal Antibody Recognizing Light-Dependent Structural Change

• Published in: Zoological Science Vol. 19; no. 6; pp. 651 - 659
• Main Authors: Takao, Masashi, Iwasa, Tatsuo, Yamamoto, Hiroaki, Takeuchi, Takuji, Tokunaga, Fumio
• Format: Journal Article
• Language: English; Japanese
• Published: Japan Zoological Society of Japan 01.06.2002; UniBio Press
• Subjects: Amino Acid Sequence; Animals; Antibodies, Monoclonal - immunology; Antibody Specificity; Astacoidea; Binding Sites, Antibody; Cattle; Concanavalin A - immunology; Conserved Sequence; epitope; Epitopes - chemistry; Epitopes - immunology; Immunoblotting; Kinetics; Light; Molecular Sequence Data; monoclonal antibody; Octopodiformes; Original s; Protein Conformation - radiation effects; rhodopsin; Rhodopsin - chemistry; Rhodopsin - immunology; Solubility; structural change; Swine
• ISSN: 0289-0003
• DOI: 10.2108/zsj.19.651

The antigenic structure of the bovine rhodopsin molecule was investigated by using a bovine rhodopsin-specific monoclonal antibody designated Rh29. 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. Rh29 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.