Labeling of sulfhydryl groups in intact mammalian cells with coumarins.
4-(Bromomethyl)-6,7-dimethoxy-2-oxo-2H-benzopyran (1) and its 3-[125I]iodo derivative (2) were reacted with sulfhydryl groups of proteins in viable Chinese hamster V79 lung fibroblasts. Pretreatment of cells with dithiothreitol (DTT) as the reducing agent was necessary to give detectable levels of fluorescence. In the absence of DTT, V79 cells incorporated about 1 x 10(-3) pCi/cell of the radioiodinated, no-carrier-added derivative; following incubation of cells with 320 microM DTT the uptake was nearly doubled (1.87 x 10(-3) pCi/cell).
Baranowska-Kortylewicz
J
J
Department of Radiology, Harvard Medical School, Boston, Massachusetts 02115.
Kassis
A I
AI
D000818:Animals; D002478:Cells, Cultured; D003374:Coumarins / Q000737:chemistry*; D006224:Cricetinae; D003412:Cricetulus; D004229:Dithiothreitol / Q000494:pharmacology; D005347:Fibroblasts / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry*; D008856:Microscopy, Fluorescence; D013438:Sulfhydryl Compounds / Q000737:chemistry*
D003374:Coumarins; D005456:Fluorescent Dyes; D013438:Sulfhydryl Compounds; D004229:Dithiothreitol