✦ LIBER ✦

Accessibility of the carbohydrate moiety of membrane-bound rhodopsin to enzymatic and chemical modification

✍ Scribed by Shaper, Joel H. ;Stryer, Lubert

Book ID: 102926481
Publisher: Wiley (John Wiley & Sons)
Year: 1977
Tongue: English
Weight: 527 KB
Volume: 6
Category: Article
ISSN: 0091-7419
DOI: 10.1002/jss.400060302

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✦ Synopsis

Abstract

Galactose was specifically inserted into the carbohydrate moiety of rhodopsin by incubating retinal disk membranes with UDP‐galactose: N‐acetylglucosamine galactosyltransferase. The stoichiometry of labeling ranged from 1.2 to 1.8 (average = 1.5) residues of galactose per molecule of rhodopsin, indicating that some or all of the oligosaccharide chains of membrane‐bound rhodopsin are readily accessible to enzymatic modification. These modified membranes were treated with galactose oxidase to generate an aldehyde at the C‐6 position of the inserted galactose units. The enzymatically‐oxidized membranes were then reacted with dansyl hydrazide to yield a fluorescent hydrazone which is sufficiently stable to permit spectroscopic analysis. This procedure for the specific attachment of a spectroscopic probe should be applicable to a wide variety of membrane glycoproteins.

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