The membrane association and dissociation of human glyceraldehyde‐3‐phosphate dehydrogenase under various conditions of hemolysis Immunochemical evidence for the lack of binding under cellular conditions

Maretzki, D; Groth, J; Tsamaloukas, Antonis; Gründel, Marianne; Krüger, Sven E.; Rapoport, S.

doi:10.1016/0014-5793(74)80022-0

Cited by 30 publications

(3 citation statements)

References 13 publications

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“…Exceptions are hexokinase (93-96% bound), phosphoglucose isomerase (8-10% bound) and glutathione reductase (26% bound). These data accord with the concept that most enzymes are free under physiological conditions (Maretzki et al, 1974) whereas some are membrane bound in hypotonic media (Solti & Friedrich, 1976;Schrier, 1977). Following pestle homogenization and density centrifugation, Holmsen et al (1969a) found 50% of platelet hexokinase activity freely soluble and the remainder in the mitochondria fraction.…”

Section: Sc U S Si 0 Nsupporting

confidence: 84%

Rapid Separation of Cytosol and Particle Fraction of Human Platelets by Digitonin‐induced Cell Damage

Akkerman¹,

Ebberink²,

Lips³

et al. 1980

Br J Haematol

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Current cell disruption and fractionation techniques are time consuming and unsuitable for metabolic studies. We have developed a rapid method for platelets in which separation of cytosol and particle fraction is obtained within 50 s. Isolated platelet suspensions were incubated with low concentrations of digitonin followed by separation of soluble and particle fraction by centrifugation through a phthalate layer. Cell disruption was 90.1+/-4.2% (mean+/-SD, n=18; lactate dehydrogenase leakage). Contamination of granules: acid hydrolase vesicles 16.2+/-3.6% (n=18, beta-N-acetylglucosaminidase), dense granules 7--9% (n=3, 14C-serotonin), mitochondrial matrix 0.6+/-0.1% (n=18, glutamate dehydrogenase). Low concentrations of digitonin did not affect sialic acid content, nucleoside diphosphate kinase and phosphodiesterase activity in isolated membranes. The method showed that most enzymes of glycolysis and hexose monophosphate shunt were localized in the cytosol except for hexokinase (96% particle bound), phosphoglucose isomerase (10% bound) and glutathion reductase (26% bound). About half the total ATP+ADP and most glycolytic intermediates were found partly particle bound, especially fructose 1,6-diphosphate (40% bound). The data suggest that in platelets glycolysis occurs in different cell compartments.

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Section: Sc U S Si 0 Nsupporting

confidence: 84%

Rapid Separation of Cytosol and Particle Fraction of Human Platelets by Digitonin‐induced Cell Damage

Akkerman¹,

Ebberink²,

Lips³

et al. 1980

Br J Haematol

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“…When bound to band 3, the enzymes' activity is inhibited [8,10,111. The binding constants strongly decrease with increasing ionic strength, so that at physiological ionic strength virtually no binding of the enzymes can be observed in vitro [6,7,[12][13][14]. Based on the latter finding, the biological relevance of the band-3 -enzyme associations has been repeatedly questioned [13 - the glycolytic enzymes present in the intact erythrocyte is, in fact, bound to band 3 [18-201. In particular, this view is strongly supported by the experiments of Rogalski et al, in which the distribution of glyceraldehyde-3-phosphate dehydrogenase in fixed, intact human erythrocytes was studied by immunolabelling [20].…”

mentioning

confidence: 99%

Associations Between Erythrocyte Band 3 Protein and Aldolase in Detergent Solution

Huber

Bäumert

Spatz-Kümbel

et al. 1996

European Journal of Biochemistry

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The cytoplasmic domain of band 3, the predominant polypeptide of the erythrocyte membrane, represents a binding site for certain glycolytic enzymes. We have studied the association between human band 3 protein and aldolase, in order to clarify the role of the different band 3 oligomers as ligand binding sites. The experiments were performed on mixtures of solubilized band 3 and aldolase in solutions of a nonionic detergent, nonaethyleneglycol lauryl ether. The main technique applied was sedimentation equilibrium analysis in an analytical ultracentrifuge. In addition, nonequilibrium centrifugation techniques were used. To facilitate the evaluations, the aldolase was labelled with a dye. The following results were obtained. (1) With unmodified band 3, aldolase is bound exclusively or at least predominantly to the band 3 tetramer (but not to monomers or dimers). (2) The band 3 tetramer can bind up to four aldolase tetramers. (3) The band 3 tetramer/aldolase complex is unstable on the time scale of the techniques used.(4) Stable band 3 dimers (stabilized either covalently or noncovalently) can also associate with aldolase and can bind up to two aldolase tetramers.The results described, together with those reported previously, point at a prominent role of the band 3 tetramer in ligand binding.

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“…It was also found that addition of Triton X-100 to the liposome-enzyme complex abolishes the effects of adsorption on Km for glyceraldehyde 3--phosphate. This nonionic detergent does not directly influence ionic interactions between lipids and proteins (Mazia & Ruby, 1968;Maretzki et al, 1974;Hallam & Wrigglesworth, 1976) but would disrupt thelow surface curvature of the liposome biiayer to form mixed mioelles of surfactant and phospholipid of smaller radius and higher surface curvature (Yedgar et al,. The results would therefore suggest that modification of enzyme activity by adsorption to a charged surface is a function of the surface area and curvature of the adsorbing surface.…”

Section: Discussionmentioning

confidence: 99%

Modification of glyceraldehyde 3-phosphate dehydrogenase activity by adsorption on phospholipid vesicles

Wooster¹,

Wrigglesworth²

1976

Biochemical Journal

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1. The adsorption of [14C]carboxymethylated glyceraldehyde 3-phosphate dehydrogenase to negatively charged liposomes of phsphatidic acid/phosphatidylcholine (3:7, w/w) was investigated. The apparent association constant at I/2 = 60, pH 7.6, was 0.4 × 10(6)M-1. Adsorption decreased as ionic strength and pH were increased. 2. In the presence of negatively charged liposomes, the Km value for glyceraldehyde 3-phosphate of glyceraldehyde 3-phosphate dehydrogenase was increased and Vmax. decreased. In the presence of positively charged liposomes, the Km value for glyceraldehyde 3-phosphate decreased and there was no significant change in Vmax. Addition of Triton X-100 abolished the effect of both positively and negatively charged liposomes on the kinetic properties of the enzyme.

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The membrane association and dissociation of human glyceraldehyde‐3‐phosphate dehydrogenase under various conditions of hemolysis Immunochemical evidence for the lack of binding under cellular conditions

Cited by 30 publications

References 13 publications

Rapid Separation of Cytosol and Particle Fraction of Human Platelets by Digitonin‐induced Cell Damage

Rapid Separation of Cytosol and Particle Fraction of Human Platelets by Digitonin‐induced Cell Damage

Associations Between Erythrocyte Band 3 Protein and Aldolase in Detergent Solution

Modification of glyceraldehyde 3-phosphate dehydrogenase activity by adsorption on phospholipid vesicles

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