<i>N</i>‐Polymethylenecarboxymaleimides—a new class of probes for membrane sulphydryl groups

Griffiths, David G.; Partis, Michael D.; Sharp, Russell N.; Beechey, R. B.

doi:10.1016/0014-5793(81)80615-1

Cited by 28 publications

(11 citation statements)

References 14 publications

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“…The working hypothesis was that a membrane protein involved in permeabilization contains a critical disulfide bond susceptible to reduction by dithiothreitol and reoxidation by oxidized glutathione. Strong support for this hypothesis resulted from experiments with glutathione maleimide, which has been shown to be an impermeant alkylating agent, unable to cross the plasma membrane [1,8,37]. This compound rendered the dithiothreitoMnduced inhibition of permeabilization insensitive to reversal by oxidized glutathione (G.…”

Section: Studies Concerned With the Mechanism Of Atp-dependent Permeamentioning

confidence: 86%

Permeabilization of transformed cells in culture by external ATP

1985

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Section: Studies Concerned With the Mechanism Of Atp-dependent Permeamentioning

confidence: 86%

Permeabilization of transformed cells in culture by external ATP

1985

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“…Arum maculatum inflorescences, in the stage of development as near as possible to that designated y (James & Beevers, 1950), were collected from local woodlands and used the same day. Silicone oils (AR200 and AR20) and UK5099 were gifts from Wacker-Chemie, Munich, W. Germany, and Pfizer (U.K.), Sandwich, Kent, U.K., respectively, and the Npolymethylenecarboxymaleimides were synthesized by Dr. D. G. Griffiths at the University of Canterbury (Griffiths et al, 1981). CHCA was obtained from Aldrich [I-14C]pyruvate, [14C]sorbitol and 3H20, were purchased from Amersham International, Amersham, Bucks, U.K., and all other chemicals from Sigma Chemical Co., Poole, Dorset, U.K., or BDH Chemicals, Chadwell Heath, Essex, U.K.…”

Section: Experimental Materialsmentioning

confidence: 99%

Pyruvate transport by thermogenic-tissue mitochondria

Proudlove

Beechey

Moore

1987

Biochemical Journal

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1. Mitochondria isolated from the thermogenic spadices of Arum maculatum and Sauromatum guttatum plants oxidized external NADH, succinate, citrate, malate, 2-oxoglutarate and pyruvate without the need to add exogenous cofactors. 2. Oxidation of substrates was virtually all via the alternative oxidase, the cytochrome pathway constituting only 10-20% of the total activity, depending on the stage of spadix development. 3. During later stages of spadix development, pyruvate oxidation was enhanced by the addition of aspartate. This was caused by acetyl-CoA condensing with oxaloacetate, produced from pyruvate/aspartate transamination, and so decreasing feedback inhibition of pyruvate dehydrogenase. 4. Pyruvate oxidation was inhibited by the long-chain acid maleimides AM5-11, but not by those with shorter polymethylene side groups, AM1-4. 5. The alpha-cyanocinnamate derivatives UK5099 [alpha-cyano-beta-(1-phenylindol-3-yl)acrylate] and CHCA [alpha-cyano-4-hydroxycinnamate] inhibited pyruvate-dependent O2 consumption and the carrier-mediated uptake of pyruvate across the mitochondrial inner membrane. Characteristics of non-competitive inhibition were observed for CHCA, whereas for UK5099 the results were more complex, suggesting a very low rate of dissociation of the inhibitor-carrier complex. 6. A comparison of the values of Vmax. and Km for oxidation and transport suggested that it was the latter which controls the overall rate of pyruvate oxidation by mitochondria from both tissues.

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“…A number of reports indicate that the acid maleimides are also impermeable to mitochondrial membranes while N E M is permeable (Moore et al, 1984;Griffiths et al, 1983Griffiths et al, , 1981. Succinate dehydrogenase is accessible from the inner side of the mitochondrial membrane.…”

Section: D~s C U S S R O~mentioning

confidence: 99%

“…N o difference was observed in the pattern of inhibition by the GSM preparation subjected to this extra purification step. NPolymethylenecarboxymaleimides (AM2, AM3, AM5, and AM10) were synthesized as indicated by Griffiths et al (1981). …”

Section: Bacterial Strain and Culture Conditions S Typhimuriummentioning

confidence: 99%

Evidence for the existence of a channel in the glucose-specific carrier EIIGlc of the Salmonella typhimurium phosphoenolpyruvate-dependent phosphotransferase system

Robillard¹,

Beechey²

1986

Biochemistry

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The effect of membrane-impermeable sulfhydryl reagents on glucose-specific enzyme I1 ( E I P ) activity has been studied in Salmonella typhimurium whole cells and in properly sealed inverted cytoplasmic membrane vesicles. Glutathione N-hexylmaleimide and N-polymethylenecarboxymaleimides inactivate methyl a-D-glucopyranoside (a-MeGlc) transport and phosphorylation in whole cell preparations at a dithiol that can be protected by oxidizing reagents, trivalent arsenicals, or phosphorylation of EIIGIC. Accessibility to this activity-linked site is restricted to small apolar reagents or to polar reagents with a hydrophobic spacer between the polar group and the reactive maleimide moiety. These same reagents inactivate a-MeGlc phosphorylation in inverted cytoplasmic membrane vesicles. Inhibition results from reaction at a dithiol that can be protected by nonpermeant mercurials, oxidants, and arsenicals as well as by phosphorylation of EII. The characteristics of this site are virtually identical with those of the activity-linked dithiol elucidated in intact cells. No evidence could be found for a second activity-linked site on the other side of the membrane when the permeable reagent A-ethylmaleimide was used. Since only one activity-linked dithiol can be detected

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N‐Polymethylenecarboxymaleimides—a new class of probes for membrane sulphydryl groups

Cited by 28 publications

References 14 publications

Permeabilization of transformed cells in culture by external ATP

Permeabilization of transformed cells in culture by external ATP

Pyruvate transport by thermogenic-tissue mitochondria

Evidence for the existence of a channel in the glucose-specific carrier EIIGlc of the Salmonella typhimurium phosphoenolpyruvate-dependent phosphotransferase system

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