title: Characterisation of 5'-AMP-activated protein kinase in human liver using specific peptide substrates and the effects of 5'-AMP analogues on enzyme activity.
abstract: A specific peptide (SAMS peptide) phosphorylation assay has previously been used to measure and subsequently purify rat liver 5'-AMP-activated protein kinase (AMPK). In this report, we show that this peptide and a peptide based on the sequence surrounding the site phosphorylated on 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase by AMPK (HMG peptide) can be used to measure human liver AMPK. Our data demonstrate that both human and rat AMPKs have a higher affinity for the HMG peptide compared to the SAMS peptide. We have used these peptide phosphorylation assays to identify novel activators of AMPK.
authors.affiliation: Cardiovascular and Metabolism Research Department, ZENECA Pharmaceuticals, Alderley Park, Cheshire, U.K.
authors.forename: J E
authors.initials: JE
authors.lastname: Sullivan
authors.forename: F
authors.initials: F
authors.lastname: Carey
authors.forename: D
authors.initials: D
authors.lastname: Carling
authors.forename: R K
authors.initials: RK
authors.lastname: Beri
mesh_terms: D055372:AMP-Activated Protein Kinases; D000227:Adenine Nucleotides / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D004789:Enzyme Activation / Q000187:drug effects; D006801:Humans; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D011494:Protein Kinases / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases; D051381:Rats; D013329:Structure-Activity Relationship
chemical_list: D000227:Adenine Nucleotides; D009097:Multienzyme Complexes; D010455:Peptides; D011494:Protein Kinases; D017346:Protein Serine-Threonine Kinases; D055372:AMP-Activated Protein Kinases