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PMID:10698692

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Citation

Cheung, PC, Salt, IP, Davies, SP, Hardie, DG and Carling, D (2000) Characterization of AMP-activated protein kinase gamma-subunit isoforms and their role in AMP binding. Biochem. J. 346 Pt 3:659-69

Abstract

The AMP-activated protein kinase (AMPK) cascade plays an important role in the regulation of energy homeostasis within the cell. AMPK is a heterotrimer composed of a catalytic subunit (alpha) and two regulatory subunits (beta and gamma). We have isolated and characterized two isoforms of the gamma subunit, termed gamma2 and gamma3. Both gamma2 (569 amino acids) and gamma3 (492 amino acids) have a long N-terminal domain which is not present in the previously characterized isoform, gamma1. As with gamma1, mRNA encoding gamma2 is widely expressed in human tissues, whereas significant expression of gamma3 mRNA was only detected in skeletal muscle. Using isoform-specific antibodies, we determined the AMPK activity associated with the different gamma isoforms in a number of rat tissues. In most tissues examined more than 80% of total AMPK activity was associated with the gamma1 isoform, with the remaining activity being accounted for mainly by the gamma2 isoform. Exceptions to this were testis and, more notably, brain where all three isoforms contributed approximately equally to activity. There was no evidence for any selective association between the alpha1 and alpha2isoforms and the various gamma isoforms. However, the AMP-dependence of the kinase complex is markedly affected by the identity of the gamma isoform present, with gamma2-containing complexes having the greatest AMP-dependence, gamma3 the lowest, and gamma1 having an intermediate effect. Labelling studies, using the reactive AMP analogue 8-azido-[(32)P]AMP, indicate that the gamma subunit may participate directly in the binding of AMP within the complex.

Links

PubMed PMC1220898

Keywords

AMP-Activated Protein Kinases; Adenosine Monophosphate/metabolism; Amino Acid Sequence; Animals; Base Sequence; Blotting, Northern; Cloning, Molecular; DNA, Complementary; Humans; Male; Molecular Sequence Data; Precipitin Tests; Protein Binding; Protein Isoforms/genetics; Protein Isoforms/metabolism; Protein Kinases/genetics; Protein Kinases/metabolism; Rats; Sequence Homology, Amino Acid

Significance

Annotations

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

RAT:A0A140UHX4

part_of

GO:0031588: nucleotide-activated protein kinase complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:A0A140UHX4

contributes_to

GO:0004679: AMP-activated protein kinase activity

ECO:0000315: mutant phenotype evidence used in manual assertion

F

Seeded From UniProt

complete

RAT:D4A7E4

part_of

GO:0031588: nucleotide-activated protein kinase complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:D4A7E4

contributes_to

GO:0004679: AMP-activated protein kinase activity

ECO:0000315: mutant phenotype evidence used in manual assertion

F

Seeded From UniProt

complete

HUMAN:AAKG1

enables

GO:0019901: protein kinase binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

RAT:AAKG1

enables

GO:0016208: AMP binding

ECO:0000315: mutant phenotype evidence used in manual assertion

F

Seeded From UniProt

complete

RAT:AAKG1

contributes_to

GO:0004679: AMP-activated protein kinase activity

ECO:0000315: mutant phenotype evidence used in manual assertion

F

Seeded From UniProt

complete

RAT:AAPK2

enables

GO:0004679: AMP-activated protein kinase activity

ECO:0000315: mutant phenotype evidence used in manual assertion

F

Seeded From UniProt

complete

RAT:Q6V7V4

part_of

GO:0031588: nucleotide-activated protein kinase complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:Q6V7V4

contributes_to

GO:0004679: AMP-activated protein kinase activity

ECO:0000315: mutant phenotype evidence used in manual assertion

F

Seeded From UniProt

complete

HUMAN:AAKG3

enables

GO:0004679: AMP-activated protein kinase activity

ECO:0000304: author statement supported by traceable reference used in manual assertion

F

Seeded From UniProt

complete

HUMAN:AAKG3

involved_in

GO:0035556: intracellular signal transduction

ECO:0000304: author statement supported by traceable reference used in manual assertion

P

Seeded From UniProt

complete

HUMAN:AAKG3

enables

GO:0019901: protein kinase binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

RAT:AAPK1

enables

GO:0004679: AMP-activated protein kinase activity

ECO:0000315: mutant phenotype evidence used in manual assertion

F

Seeded From UniProt

complete

HUMAN:AAKG2

enables

GO:0019901: protein kinase binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete


See also

References

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