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PMID:10913113

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Citation

Muñoz-Alonso, MJ, Guillemain, G, Kassis, N, Girard, J, Burnol, AF and Leturque, A (2000) A novel cytosolic dual specificity phosphatase, interacting with glucokinase, increases glucose phosphorylation rate. J. Biol. Chem. 275:32406-12

Abstract

A novel protein was cloned from a rat liver cDNA library by interaction with the liver glucokinase. This protein contained 339 residues and possessed a canonical consensus sequence for a dual specificity phosphatase. The recombinant protein was able to dephosphorylate phosphotyrosyl and phosphoseryl/threonyl substrates. We called this protein the glucokinase-associated phosphatase (GKAP). The GKAP partially dephosphorylated the recombinant glucokinase previously phosphorylated, in vitro, by protein kinase A. The GKAP fused with green fluorescent protein was located in the cytosol, where glucokinase phosphorylates glucose, and not in the nucleus where the glucokinase is retained inactive by the glucokinase regulatory protein. More importantly, the GKAP accelerated the glucokinase activity in a dose-dependent manner and with a stoichiometry compatible with a physiological mechanism. This strongly suggested that the interaction between GKAP and glucokinase had a functional significance. The cloning of this novel protein with a dual specificity phosphatase activity allows the description of a possible new regulatory step in controlling the glycolysis flux.

Links

PubMed Online version:10.1074/jbc.M000841200

Keywords

Amino Acid Sequence; Animals; Base Sequence; Cell Nucleus/enzymology; Cloning, Molecular; Cytosol/enzymology; DNA, Complementary/genetics; Dual-Specificity Phosphatases; Glucokinase/metabolism; Glucose/metabolism; Glutathione Transferase/metabolism; Green Fluorescent Proteins; Kinetics; Luminescent Proteins/metabolism; Molecular Sequence Data; Phosphoprotein Phosphatases/chemistry; Phosphoprotein Phosphatases/genetics; Phosphoprotein Phosphatases/metabolism; Phosphorylation; Rats; Recombinant Fusion Proteins/metabolism; Sequence Alignment; Sequence Homology, Amino Acid; Substrate Specificity

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

RAT:DUS12

enables

GO:0019900: kinase binding

ECO:0000353: physical interaction evidence used in manual assertion

RGD:2670

F

Seeded From UniProt

complete

RAT:DUS12

involved_in

GO:0033133: positive regulation of glucokinase activity

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

RAT:DUS12

located_in

GO:0005737: cytoplasm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:DUS12

enables

GO:0004721: phosphoprotein phosphatase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

RAT:DUS12

involved_in

GO:0006470: protein dephosphorylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

RAT:HXK4

enables

GO:0019903: protein phosphatase binding

ECO:0000353: physical interaction evidence used in manual assertion

RGD:68375

F

Seeded From UniProt

complete


See also

References

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