Study of the orientation of retinal in bovine rhodopsin: the use of a photoactivatable retinal analog

Conference · Fri May 01 00:00:00 EDT 1987 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:6180117

Nakayama, T

Rhodopsin is the major transmembrane protein in the photoreceptor cells of vertebrate and invertebrate retina. Bovine rhodopsin consists of a polypeptide chain of 348 amino acids of known sequence in which the chromophore, 11-cis-retinal, is linked to Lys-296 as a Schiff base. To investigate the orientation of retinal in the protein and to study the interactions between retinal and the protein, the authors have developed a crosslinking approach using a /sup 3/H-labeled photoactivatable analog of retinal. Bleached rhodopsin in rod outer segments was reconstituted with the analog to give a pigment with lambda/sub max/ at 460nm. Reduction of the Schiff base with borane dimenthylamine, followed by degradation with CNBr and sequencing of the radioactive fragment showed that the analog is attached to Lys-296, as in the native rhodopsin. Further, the reconstitute protein after photolysis was phosphorylated by rhodopsin kinase. Photolysis of the reconstituted pigment at -15/sup 0/C resulted in crosslinking of the analog to the opsin to the extent of 30% as analyzed by SDS electrophoresis. The site(s) of crosslinking in the protein are under investigation.

• Research Organization: Massachusetts Institute of Technology, Cambridge
• OSTI ID: 6180117
• Report Number(s): CONF-870644-; TRN: 87-033990
• Journal Information: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 46:6; Conference: 78. annual meeting of the American Society of Biological Chemists conference, Philadelphia, PA, USA, 7 Jun 1987
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
CELL MEMBRANES
STRUCTURE-ACTIVITY RELATIONSHIPS
RETINOIC ACID
CROSS-LINKING
RHODOPSIN
MOLECULAR STRUCTURE
AMINO ACID SEQUENCE
CATTLE
ELECTROPHORESIS
PHOSPHORYLATION
RETINA
SCHIFF BASES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ANIMALS
BODY
BODY AREAS
CARBOXYLIC ACID ESTERS
CELL CONSTITUENTS
CHEMICAL REACTIONS
DOMESTIC ANIMALS
ESTERS
EYES
FACE
HEAD
IMINES
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MAMMALS
MEMBRANES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANS
PIGMENTS
POLYMERIZATION
PROTEINS
RUMINANTS
SENSE ORGANS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques