Effect of electrical stimulation <i>post mortem</i> of bovine muscle on the binding of glycolytic enzymes. Functional and structural implications

Clarke, Frank M.; Shaw, F.D.; Morton, Don

doi:10.1042/bj1860105

Cited by 84 publications

(35 citation statements)

References 35 publications

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“…In such a dual role, glycolytic enzymes could be integrators of cytoplasmic structure and function, by binding to actin as well as to substrate. This exciting idea is supported by evidence obtained in vitro that (a) the actin-geUing activity of aldolase is inhibited by its substrate fructose 1,6-diphosphate (FDP; 18), (b) the kinetics of aldolase activity are modified by the actin-containing filaments of skeletal muscle (74), and (c) the actin filaments of muscle tissue perturbed by electrical stimulation (19) or anoxia (20) bind a substantially higher percentage of aldolase than do those of unperturbed muscle. In view of the above evidence, we agree with Clarke et al (21) that aldolase-actin interactions cannot be prejudged as "non-specific" (24).…”

supporting

confidence: 48%

“…The uniform distribution of Rh-aldolase we observed in vivo initally seemed paradoxical, in light of data demonstrating that aldolase hinds to F-actin (7,15,16), skeletal muscle homogenates (19), and fetal brain homogehates (17) with affinities greater than or equal to those of other glycolytic enzymes. However, the spatial differences we measured in the mobile fraction and diffusion coefficient of Rh-aldolase indicate that a sensitive equilibrium may regulate the actin-binding activity of aldolase in vivo.…”

Section: Discussionmentioning

confidence: 99%

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Aldolase exists in both the fluid and solid phases of cytoplasm [published erratum appears in J Cell Biol 1988 Dec;107(6 Pt 1):following 2463]

Pagliaro

Taylor

1988

The Journal of Cell Biology

115

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Abstract. We have prepared a functional fluorescent analogue of the glycolytic enzyme aldolase (rhodamine [Rh]-aldolase), using the succinimidyl ester of carboxytetramethyl-rhodamine. Fluorescence redistribution after photobleaching measurements of the diffusion coefficient of Rh-aldolase in aqueous solutions gave a value of 4.7 × 10 -7 cm2/s, and no immobile fraction. In the presence of filamentous actin, there was a 4.5-fold reduction in diffusion coefficient, as well as a 36% immobile fraction, demonstrating binding of Rh-aldolase to actin. However, in the presence of a 100-fold molar excess of its substrate, fructose 1,6-diphosphate, both the mobile fraction and diffusion coefficient of Rh-aldolase returned to control levels, indicating competition between substrate binding and actin cross-linking. When Rh-aldolase was microinjected into Swiss 3T3 cells, a relatively uniform intracellular distribution of fluorescence was observed. However, there were significant spatial differences in the in vivo diffusion coefficient and mobile fraction of Rh-aldolase measured with fluorescence redistribution after photobleaching. In the perinuclear region, we measured an apparent cytoplasmic diffusion coefficient of 1.1 × 10 -7 cm2/s with a 23% immobile fraction; while measurements in the cell periphery gave a value of 5.7 × 10 -g cm2/s, with no immobile fraction. Ratio imaging of Rh-aldolase and FITCdextran indicated that FITC-dextran was relatively excluded from stress fiber domains. We interpret these data as evidence for the partitioning of aldolase between a soluble fraction in the fluid phase and a fraction associated with the solid phase of cytoplasm. The partitioning of aldolase and other glycolytic enzymes between the fluid and solid phases of cytoplasm could play a fundamental role in the control of glycolysis, the organization of cytoplasm, and cell motility. The concepts and experimental approaches described in this study can be applied to other cellular biochemical processes.TIaOUGH glycolytic metabolism is well-defined at the biochemical level, the dynamic and spatial organization of glycolysis in living cells has not been characterized to date. After the elucidation of mitochondrial structure and function 35 years ago (28) it was generally assumed that a corresponding organelle for glycolysis must exist. Efforts to isolate and to characterize such an organelle were unsuccessful, leading de Duve (26) to conclude that the longsought"glycolytic particle" did not exist. As a result, the current concept of in vivo glycolytic metabolism is based on an assumption of dilute solution chemistry, due largely to the absence of evidence for a discrete glycolytic organelle.There has been evidence for over 20 years that some glycoiytic enzymes bind to structural proteins, particularly actin.

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supporting

confidence: 48%

Section: Discussionmentioning

confidence: 99%

Aldolase exists in both the fluid and solid phases of cytoplasm [published erratum appears in J Cell Biol 1988 Dec;107(6 Pt 1):following 2463]

Pagliaro

Taylor

1988

The Journal of Cell Biology

115

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“…A redistribution of glycolytic enzymes between bound and soluble fractions has been noted in several systems as a response to stimuli that change glycolytic flux (e.g., exercise, anoxia, hypoxia) (26)(27)(28), including trout white muscle (12,29). In general, an increase in enzyme binding correlates with an increase in glycolytic flux and vice versa.…”

Section: Introductionmentioning

confidence: 99%

Influence of exercise on the activity and the distribution between free and bound forms of glycolytic and associated enzymes in tissues of horse mackerel

Lushchak

Bagnyukova

Storey

et al. 2001

Braz J Med Biol Res

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The effects of short-term burst (5 min at 1.8 m/s) swimming and longterm cruiser (60 min at 1.2 m/s) swimming on maximal enzyme activities and enzyme distribution between free and bound states were assessed for nine glycolytic and associated enzymes in tissues of horse mackerel, Trachurus mediterraneus ponticus. The effects of exercise were greatest in white muscle. The activities of phosphofructokinase (PFK), pyruvate kinase (PK), fructose-1,6-bisphosphatase (FBPase), and phosphoglucomutase (PGM) all decreased to 47, 37, 37 and 67%, respectively, during 60-min exercise and all enzymes except phosphoglucoisomerase (PGI) and PGM showed a change in the extent of binding to subcellular particulate fractions during exercise. In red muscle, exercise affected the activities of PGI, FBPase, PFK, and lactate dehydrogenase (LDH) and altered percent binding of only PK and LDH. In liver, exercise increased the PK activity 2.3-fold and reduced PGI 1.7-fold only after 5 min of exercise but altered the percent binding of seven enzymes. Fewer effects were seen in brain, with changes in the activities of aldolase and PGM and in percent binding of hexokinase, PFK and PK. Changes in enzyme activities and in binding interactions with subcellular particulate matter appear to support the altered demands of tissue energy metabolism during exercise.

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“…Reduced enzyme binding was also associated with anoxia-induced metabolic rate depression in the marine gastropod, Busycotypus canaliculatum, with the most pronounced changes seen for HK and ALD (percentage of bound enzyme decreased from 40-45 % to 9-10 % in anoxia), similar to the situation in O. lactea (Plaxton and Storey, 1986). The opposite response, increased binding of enzymes to the particulate fraction, has been documented in several systems during glycolytic rate activation, including ischemia in rat and sheep hearts (Clarke et al 1984), contraction in skeletal muscle (Clarke et al 1980) and burst exercise in trout white muscle (Brooks and Storey, 19886). In all these instances, decreased enzyme binding correlated with a decreased glycolytic rate, whereas increased enzyme binding correlated with an increased glycolytic rate.…”

Section: Discussionmentioning

confidence: 80%

“…The method of Clarke et al (1980) offers a rapid and reproducible technique for measuring enzyme binding to the particulate fraction of the cell. In this method, the tissue is homogenized in a high-sucrose buffer that stabilizes enzyme-particle associations during the homogenization and centrifugation steps.…”

Section: Resultsmentioning

confidence: 99%

Glycolytic enzyme binding and metabolic control in anaerobiosis

Plaxton

Storey

1986

J Comp Physiol B

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SummaryThe mechanisms controlling glycolytic rate were examined in foot muscle of the terrestrial snail Otala lactea (Miiller) (Pulmonata, Helicidae), during short and long periods of estivation and anoxia. Binding associations between glycolytic enzymes and the particulate fraction of the cell were assessed in both states. The percentage of enzyme activity bound to particulate matter decreased significantly over the short term (4 days estivation and 14.5 h anoxia); significant changes were seen for hexokinase (HK), phosphofructokinase (PFK), aldolase and lactate dehydrogenase (LDH) in estivation and, for these enzymes plus triosephosphate isomerase and pyruvate kinase (PK), in anoxia. Over the longer term in estivation (22 days) and anoxia (45 h), enzyme binding returned to control values. Tissue content of fructose-2,6-bisphosphate, a potent phosphofructokinase activator, decreased under all experimental conditions. Total glycogen phosphorylase activity decreased during short-term anoxia (14.5 h) and during long-term estivation (22 days), but the percentage of the active a form decreased significantly during anoxia only. Significant changes in the maximal activities of several enzymes were observed during both estivation and anoxia. Decreases in the maximal activity of HK, PFK, glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase (PGK) and LDH were observed during long-term estivation. Increases in PGK and PK maximal activity in short-term anoxia and aldolase and PGK in long-term anoxia were also observed. These results suggest that changes in glycolytic enzyme binding may be part of an immediate mechanism used to cause a rapid decrease in glycolytic flux and initiate glycolytic rate depression, which also includes a reduction of fructose-2,6-bisphosphate content and decreased glycogen phosphorylase activity. In the long term, however, control of snail glycolytic rate is reorganized, so that enzyme binding associations revert to the control values. In the long term, then, control is mediated by lower fructose-2,6-bisphosphate concentrations and, during estivation, also by a decrease in maximal enzyme activities.

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Effect of electrical stimulation post mortem of bovine muscle on the binding of glycolytic enzymes. Functional and structural implications

Cited by 84 publications

References 35 publications

Aldolase exists in both the fluid and solid phases of cytoplasm [published erratum appears in J Cell Biol 1988 Dec;107(6 Pt 1):following 2463]

Aldolase exists in both the fluid and solid phases of cytoplasm [published erratum appears in J Cell Biol 1988 Dec;107(6 Pt 1):following 2463]

Influence of exercise on the activity and the distribution between free and bound forms of glycolytic and associated enzymes in tissues of horse mackerel

Glycolytic enzyme binding and metabolic control in anaerobiosis

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