The effects of sulfhydryl modifying reagents on nonhormonal and hormonally regulated hexose transport in cultured human skin fibroblasts. 1987

R J Germinario, and F Vlachopoulou

The effects of various sulfhydryl modifying reagents on hexose transport in cultured human skin fibroblasts were studied. H2O2 was observed to have no effect on 2-deoxy-D-glucose transport in serum-starved glucose-fed cells. The elevation of hexose transport rates in cells by glucose deprivation, insulin, or serum stimulation rendered them sensitive to H2O2. Hexose transport in glucose-deprived cells was inhibited 51-55% by 1-2 mM H2O2, while hexose transport in insulin or serum-stimulated glucose-fed cells was inhibited 45% and 46%, respectively. H2O2 inhibition was blocked or reversed by 8 mM dithiothreitol. N-ethyl-maleimide (NEM), a permeant, sulfhydryl reagent, elicited effects on hexose transport similar to those effected by H2O2 (i.e., in glucose-deprived and insulin-stimulated cells, inhibition of hexose transport was 44% and 23%, respectively). Impermeant sulfhydryl reagents such as dithio(bis)nitrobenzoic acid (DTNB) and N-iodoacetyl-N'-(5-sulfo-1-naphthly-ethylenediame (1,5,-I-AEDANS) had no inhibitory effect on hexose transport under any conditions (i.e., glucose-fed, glucose-deprived, and insulin-stimulated cells). DTNB and 1,5-I-AEDANS afforded no protection from the action of H2O2 on hexose transport. The data suggest that the sensitive sites are thiol in nature and are located at an intramembrane or intracellular site and probably not exofacial.

UI MeSH Term Description Entries
D007328 Insulin A 51-amino acid pancreatic hormone that plays a major role in the regulation of glucose metabolism, directly by suppressing endogenous glucose production (GLYCOGENOLYSIS; GLUCONEOGENESIS) and indirectly by suppressing GLUCAGON secretion and LIPOLYSIS. Native insulin is a globular protein comprised of a zinc-coordinated hexamer. Each insulin monomer containing two chains, A (21 residues) and B (30 residues), linked by two disulfide bonds. Insulin is used as a drug to control insulin-dependent diabetes mellitus (DIABETES MELLITUS, TYPE 1). Iletin,Insulin A Chain,Insulin B Chain,Insulin, Regular,Novolin,Sodium Insulin,Soluble Insulin,Chain, Insulin B,Insulin, Sodium,Insulin, Soluble,Regular Insulin
D007700 Kinetics The rate dynamics in chemical or physical systems.
D009004 Monosaccharide Transport Proteins A large group of membrane transport proteins that shuttle MONOSACCHARIDES across CELL MEMBRANES. Hexose Transport Proteins,Band 4.5 Preactin,Erythrocyte Band 4.5 Protein,Glucose Transport-Inducing Protein,Hexose Transporter,4.5 Preactin, Band,Glucose Transport Inducing Protein,Preactin, Band 4.5,Proteins, Monosaccharide Transport,Transport Proteins, Hexose,Transport Proteins, Monosaccharide,Transport-Inducing Protein, Glucose
D002478 Cells, Cultured Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others. Cultured Cells,Cell, Cultured,Cultured Cell
D003837 Deoxy Sugars Sugars that in which one or more hydroxyl groups of the pyranose or furanose ring is substituted by hydrogen. Deoxy Sugar,Sugar, Deoxy,Sugars, Deoxy
D003847 Deoxyglucose 2-Deoxy-D-arabino-hexose. An antimetabolite of glucose with antiviral activity. 2-Deoxy-D-glucose,2-Deoxyglucose,2-Desoxy-D-glucose,2 Deoxy D glucose,2 Deoxyglucose,2 Desoxy D glucose
D004228 Dithionitrobenzoic Acid A standard reagent for the determination of reactive sulfhydryl groups by absorbance measurements. It is used primarily for the determination of sulfhydryl and disulfide groups in proteins. The color produced is due to the formation of a thio anion, 3-carboxyl-4-nitrothiophenolate. 5,5'-Dithiobis(2-nitrobenzoic Acid),DTNB,Ellman's Reagent,5,5'-Dithiobis(nitrobenzoate),Acid, Dithionitrobenzoic,Ellman Reagent,Ellmans Reagent,Reagent, Ellman's
D004229 Dithiothreitol A reagent commonly used in biochemical studies as a protective agent to prevent the oxidation of SH (thiol) groups and for reducing disulphides to dithiols. Cleland Reagent,Cleland's Reagent,Sputolysin,Clelands Reagent,Reagent, Cleland,Reagent, Cleland's
D005033 Ethylmaleimide A sulfhydryl reagent that is widely used in experimental biochemical studies. N-Ethylmaleimide,N Ethylmaleimide
D005347 Fibroblasts Connective tissue cells which secrete an extracellular matrix rich in collagen and other macromolecules. Fibroblast

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