Title

Mapping sites of interaction between rhodopsin and transducin using rhodopsin antipeptide antibodies

UMMS Affiliation

Department of Biochemistry

Date

5-5-1988

Document Type

Article

Subjects

Animals; *Antibodies; Binding Sites; Immunosorbent Techniques; Membrane Proteins; Rabbits; Retinal Pigments; Rhodopsin; Transducin

Disciplines

Life Sciences | Medicine and Health Sciences

Abstract

Site-directed antipeptide antibodies generated against the predicted cytoplasmic sequences of rhodopsin were used to map the binding domains for transducin, the retinal G-protein, on the photoreceptor. Antibodies against synthetic peptides corresponding to loop 3-4, loop 5-6, and the serine/threonine-rich region of the COOH terminus recognize rhodopsin by immunoblot analysis and also recognize the native protein within the membrane, allowing these probes to be used for functional studies. Rhodopsin reconstituted into phospholipid vesicles binds transducin in the light which significantly reduces the binding of antipeptide antibodies corresponding to loop 3-4 and the COOH terminus of rhodopsin. However, the binding of the antibody raised against a 14-amino-acid peptide corresponding to a sequence within loop 5-6 of rhodopsin was unaffected by the presence of transducin. These results suggest a preferential involvement of regions in or near loop 3-4 and the COOH terminus in the binding of transducin to rhodopsin. In contrast, a significant portion of loop 5-6 does not form a binding domain for the G-protein.

Rights and Permissions

Citation: J Biol Chem. 1988 May 5;263(13):6150-4.

Related Resources

Link to Article in PubMed

Journal Title

The Journal of biological chemistry

PubMed ID

3283122