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THE USE OF SUGARS AND FLUORODINITROBENZENE (FDNB) TO DIFFERENTIALLY LABEL RED CELL MEMBRANE COMPONENTS INVOLVED IN HEXOSE TRANSFERS
Author(s) -
Eady R. P.,
Widdas W. F.
Publication year - 1973
Publication title -
quarterly journal of experimental physiology and cognate medical sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.925
H-Index - 101
eISSN - 1469-445X
pISSN - 0033-5541
DOI - 10.1113/expphysiol.1973.sp002191
Subject(s) - incubation , labelling , sugar , chemistry , biochemistry , hexose , membrane , chromatography , polyacrylamide gel electrophoresis , intracellular , electrophoresis , enzyme
The possibility of differentially labelling human red cell membrane components involved in the facilitated transfer of hexoses by incubation with 1‐fluoro‐2,4‐dinitrobenzene (FDNB) irr the presence of different sugars, one with a potentiating effect on the inhibitory reaction and the other with a protective effect, has been explored. After incubation with 14 C‐FDNB, membrane proteins were subjected to SDS polyacrylamide gel disc electrophoresis and gel fractions were assessed for radioactive labelling. Early fractions from cells incubated with 2‐deoxyglucose (the potentiating sugar) carried more label than those from cells incubated with ethylidene glucose (the protecting sugar). In both types of incubations the bulk of the label was taken up nonspecifically by intracellular material and membrane proteins but this technique and a method based on the preferential uptake of D‐glucose from a D‐/L‐glucose mixture point to fractions which merit further study by methods of increased resolving power.