Biological disulfides: the third messenger? Modulation of phosphofructokinase activity by thiol/disulfide exchange.

Gilbert, Hiram F.

doi:10.1016/s0021-9258(18)33682-2

Cited by 248 publications

(25 citation statements)

References 39 publications

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“…for CHO cell uptake of the nonmetabolized sugar 3-O-methylglucose (100 ,«M) at 4 °C is 40 s]. While we cannot rule out the possibility that DTT affects the 2-deoxy-D-glucose phosphorylation step and not transport, this appears unlikely since the redox state of cytosol in viable cells is reducing in the absence of exogenous reductant (Gilbert, 1982). b, e, f) of 2 mM DTT.…”

Section: Resultsmentioning

confidence: 91%

Glucose Transporter Function Is Controlled by Transporter Oligomeric Structure. A Single, Intramolecular Disulfide Promotes GLUT1 Tetramerization

Zottola¹,

Cloherty²,

Coderre³

et al. 1995

Biochemistry

124

150

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The human erythrocyte glucose transporter is an allosteric complex of four GLUT1 proteins whose structure and substrate binding properties are stabilized by reductant-sensitive, noncovalent subunit interactions [Hebert, D. N., & Carruthers, A. (1992) J. Biol. Chem. 267, 23829-23838]. In the present study, we use biochemical and molecular approaches to isolate specific determinants of transporter oligomeric structure and transport function. When unfolded in denaturant, each subunit (GLUT1 protein) of the transporter complex exposes two sulfhydryl groups. Four additional thiol groups are accessible following subunit exposure to reductant. Assays of subunit disulfide bridge content suggest that two inaccessible sulfhydryl groups form an internal disulfide bridge. Differential alkylation/peptide mapping/N-terminal sequence analyses show that a GLUT1 carboxyl-terminal peptide (residues 232-492) contains three inaccessible sulfhydryl groups and that an N-terminal GLUT1 peptide (residues 147-261/299) contains two accessible thiols. The carboxyl-terminal peptide most likely contains the intramolecular disulfide bridge since neither its yield nor its electrophoretic mobility is altered by addition of reductant. Each GLUT1 cysteine was changed to serine by oligonucleotide-directed, in vitro mutagenesis. The resulting transport proteins were expressed in CHO cells and screened by immunofluorescence microscopy for their ability to expose tetrameric GLUT1-specific epitopes. Serine substitution at cysteine residues 133, 201, 207, and 429 does not inhibit exposure of tetrameric GLUT1-specific epitopes. Serine substitution at cysteines 347 or 421 prevents exposure of tetrameric GLUT1-specific epitopes. Hydrodynamic analysis of GLUT1/GLUT4 chimeras expressed in and subsequently solubilized from CHO cells indicates that GLUT1 residues 1-199 promote chimera dimerization and permit GLUT1/chimera heterotetramerization. This GLUT1 N-terminal domain is insufficient for chimera tetramerization which additionally requires GLUT1 residues 200-463. Extracellular reductants (dithiothreitol, beta-mercaptoethanol, or glutathione) reduce erythrocyte 3-O-methylglucose uptake by up to 15-fold. This noncompetitive inhibition of sugar uptake is reversed by the cell-impermeant, oxidized glutathione. Reductant is without effect on sugar exit from erythrocytes. Dithiothreitol doubles the cytochalasin B binding capacity of erythrocyte-resident glucose transporter, abolishes allosteric interactions between substrate binding sites on adjacent subunits, and occludes tetrameric GLUT1-specific GLUT1 epitopes in situ. CHO cell-resident GLUT1 structure and transport function are similarly affected by extracellular reductant. We conclude that each subunit of the glucose transporter contains an extracellular disulfide bridge (Cys347 and Cys421) that stabilizes transporter oligomeric structure and thereby accelerates transport function.

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Section: Resultsmentioning

confidence: 91%

Glucose Transporter Function Is Controlled by Transporter Oligomeric Structure. A Single, Intramolecular Disulfide Promotes GLUT1 Tetramerization

Zottola¹,

Cloherty²,

Coderre³

et al. 1995

Biochemistry

124

150

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“…In biological areas, the thiol-disulde exchange reaction has been found to be closely related to signal transduction, thiol protection and switching between the different conformational and functional states of enzymes. [50][51][52][53] In biomedical applications, it has been exploited extensively for constructing redoxresponsive prodrugs, and gene and drug carriers. 34,[54][55][56][57][58] For example, Meng and Zhong's group designed a facile way to prepare the disulde-linked dextran-b-poly(3-caprolactone) amphiphilic block copolymer using the thiol-disulde exchange reaction under mild conditions (Scheme 2).…”

Section: Redox-responsive Polymeric Drug Delivery Systemsmentioning

confidence: 99%

Redox-responsive polymers for drug delivery: from molecular design to applications

et al. 2014

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Glutathione has been regarded as a significant signal for distinguishing between tumor and normal tissue. Recently, reactive oxygen species have attracted much attention for their close connection with many diseases. Taking advantage of the physiological signals, redox-responsive polymeric drug carriers constitute a significant research area in the various stimuli-responsive polymers for biomedical applications. During the rapid development of redox-responsive polymers, molecular design and related synthetic methodology plays a crucial role. In this review, we discuss the reduction-and oxidationresponsive polymeric drug carriers from the view of functional groups, as well as their applications in controlled release.

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“…Increasing the intracellular concentration of GSSG by oxidative stress is positively correlated with an increase in protein-GSH mixed disulfides (15). Formation of mixed disulfides of protein thiols and GSH may affect protein structure and serve a regulatory function (16,17). For example, S-glutathiolation of Cys residues by this process has been shown to modulate the phosphatase activity of carbonic anhydrase III (18).…”

Section: Introductionmentioning

confidence: 99%

Comparative Inhibition of Yeast Glutathione Reductase by Arsenicals and Arsenothiols

Stýblo

Serves

Cullen

et al. 1997

Chem. Res. Toxicol.

269

155

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Tri(gamma-glutamylcysteinylglycinyl)trithioarsenite (AsIII(GS)3) is formed in cells and is a more potent mixed-type inhibitor of the reduction of glutathione disulfide (GSSG) by yeast glutathione (GSH) reductase than either arsenite (AsIII) or GSH. The present work examines the effects of valence and complexation of arsenicals with GSH or L-cysteine (Cys) upon potency as competitive inhibitors of the reduction of GSH disulfide (GSSG) by yeast GSH reductase. Trivalent arsenicals were more potent inhibitors than their pentavalent analogs, and methylated trivalent arsenicals were more potent inhibitors than was inorganic trivalent As. Complexation of either inorganic trivalent As or methylarsonous diiodide (CH3As(III)I2) with Cys or GSH produced inhibitors of GSH reductase that were severalfold more potent than the parent arsenicals. In contrast, dimethylarsinous iodide ((CH3)2As(III)I) was a more potent inhibitor than its complexes with either GSH or Cys. Complexes of CH3AsIII with GSH (CH3-AsIII(GS)2) or with Cys (CH3AsIII(Cys)2) were the most potent inhibitors, with Ki's of 0.009 and 0.018 mM, respectively. Inhibition of GSH reductase by arsenicals or arsenothiols was prevented by addition of meso-2,3-dimercaptosuccinic acid (DMSA) to a mixture of enzyme, GSSG, and inhibitor before addition of NADPH. DMSA added to the reaction mixture after NADPH reversed inhibition by (CH3)2As(III)I but had little effect on inhibition by CH3As(III)I2, Ch3AsIII(GS)2, CH3AsIII(Cys)2, or AsIII(GS)3. Partial redox inactivation of the enzyme with NADPH increased the inhibitory potency of CH3As(III)I2 and (CH3)2As(III)I and changed the mode of inhibition for CH3As(III)I2 from competitive to noncompetitive. The greater potency of methylated trivalent arsenicals and arsenothiols than of inorganic trivalent As suggests that biomethylation of As could yield species that inhibit reduction of GSSG and alter the redox status of cells.

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Biological disulfides: the third messenger? Modulation of phosphofructokinase activity by thiol/disulfide exchange.

Cited by 248 publications

References 39 publications

Glucose Transporter Function Is Controlled by Transporter Oligomeric Structure. A Single, Intramolecular Disulfide Promotes GLUT1 Tetramerization

Glucose Transporter Function Is Controlled by Transporter Oligomeric Structure. A Single, Intramolecular Disulfide Promotes GLUT1 Tetramerization

Redox-responsive polymers for drug delivery: from molecular design to applications

Comparative Inhibition of Yeast Glutathione Reductase by Arsenicals and Arsenothiols

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