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Labelling of the human erythrocyte glucose transporter with 3H-labelled cytochalasin B occurs via protein photoactivation
Authors:M Deziel  W Pegg  E Mack  A Rothstein  A Klip
Abstract:Irradiation of human erythrocyte membranes with 3H-labelled cytochalasin B results in specific photolabelling of the glucose transporter. The action spectrum of photolabelling has a maximum at approx. 280 nm, whereas the absorption spectrum of cytochalasin B is maximal at 210 nm. By irradiating with narrow-band-width light centered at 280 nm for 2 h, 8% of the transporters become covalently labelled and 47% of the remaining cytochalasin B-binding sites are obliterated. We conclude that photolabelling driven by narrow-bandwidth irradiation proceeds via photoactivation of an aromatic amino acid residue on the transporter molecule, and when compared to wide-bandwidth irradiation, permits more efficient incorporation of the label without causing additional photodamage to the remaining transporters.
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