Localization and membrane association of aldolase in human erythrocytes

Yeltman, Don R.; Harris, Ben G.

doi:10.1016/0003-9861(80)90272-6

Cited by 22 publications

(2 citation statements)

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“…The association of the enzyme with the membranes in situ is determined by measuring the aldolase crosslinked to the microsomes by perfusing the liver with a dilute solution of glutaraldehyde prior to homogenization. This cross-linking technique has been used to investigate the localization of aldolase in human erythrocytes [38].…”

Section: Discussionmentioning

confidence: 99%

Reversible microsomal binding of hepatic aldolase

Weiss

Zieske

Bernstein

1981

Biochimica et Biophysica Acta (BBA) - Enzymology

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Fructose-1,6-bisphosphate aldolase (D-fructose-1,6-bisphosphate D-glyceraldehyde-3-phosphate lyase, EC 4.1.2.13) partitions between the microsomes and the cytosol when a rat liver homogenate is fractionated by differential centrifugation. Gel electrophoresis and immunodiffusion indicate that the one isozyme present in the liver of the young adult rat is found in both fractions. The association of the aldolase with membranes is differentially sensitive to a variety of metabolites and inorganic salts. In the absence of cellular salts, 1 mM fructose 1,6-bisphosphate or glucose 1,6-bisphosphate elutes 50% of the enzyme from the microsomes. About 9 mM Pi or citrate is necessary to produce the same effect. With other metabolites or inorganic salts higher concentrations are required. The fraction of total enzyme which partitions with the microsomes when a homogenate is submitted to high speed centrifugation, correlates inversely with the level of fructose 1,6-bisphosphate in the supernatant solution and this concentration is higher when the tissue concentration in the homogenate is greater. The Km for fructose 1,6-bisphosphate of 3 . 10(-4) for aldolase bound to microsomes is decreased to 6 . 10(-6) M when the enzyme is dissociated from the membranes with salt. These observations appear relevant to the ongoing discussion regarding the physiological relevance of the subcellular localization of glycolytic enzymes.

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Section: Discussionmentioning

confidence: 99%

Reversible microsomal binding of hepatic aldolase

Weiss

Zieske

Bernstein

1981

Biochimica et Biophysica Acta (BBA) - Enzymology

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“…This indirect binding is referred to as enzyme-enzymeactin or piggy-back interaction. Furthermore, the glycolytic enzymes, aldolase and GAPDH, have also been shown to compete against one another for binding sites (17). Muscle actin-enzyme interactions have been shown to be electrostatic in nature, owing to their dependence on factors such as ionic strength (14,15).…”

Section: Introductionmentioning

confidence: 99%

Glycolytic Enzyme Interactions with Yeast and Skeletal Muscle F-Actin

Waingeh

Gustafson

Kozliak

et al. 2006

Biophysical Journal

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Interaction of glycolytic enzymes with F-actin is suggested to be a mechanism for compartmentation of the glycolytic pathway. Earlier work demonstrates that muscle F-actin strongly binds glycolytic enzymes, allowing for the general conclusion that "actin binds enzymes", which may be a generalized phenomenon. By taking actin from a lower form, such as yeast, which is more deviant from muscle actin than other higher animal forms, the generality of glycolytic enzyme interactions with actin and the cytoskeleton can be tested and compared with higher eukaryotes, e.g., rabbit muscle. Cosedimentation of rabbit skeletal muscle and yeast F-actin with muscle fructose-1,6-bisphosphate aldolase (aldolase) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) followed by Scatchard analysis revealed a biphasic binding, indicating high- and low-affinity domains. Muscle aldolase and GAPDH showed low-affinity for binding yeast F-actin, presumably because of fewer acidic residues at the N-terminus of yeast actin; this difference in affinity is also seen in Brownian dynamics computer simulations. Yeast GAPDH and aldolase showed low-affinity binding to yeast actin, which suggests that actin-glycolytic enzyme interactions may also occur in yeast although with lower affinity than in higher eukaryotes. The cosedimentation results were supported by viscometry results that revealed significant cross-linking at lower concentrations of rabbit muscle enzymes than yeast enzymes. Brownian dynamics simulations of yeast and muscle aldolase and GAPDH with yeast and muscle actin compared the relative association free energy. Yeast aldolase did not specifically bind to either yeast or muscle actin. Yeast GAPDH did bind to yeast actin although with a much lower affinity than when binding muscle actin. The binding of yeast enzymes to yeast actin was much less site specific and showed much lower affinities than in the case with muscle enzymes and muscle actin.

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