Posttranslational modifications of the 5'-AMP-activated protein kinase beta1 subunit.
The AMP-activated protein kinase (AMPK) consists of catalytic alpha and noncatalytic beta and gamma subunits and is responsible for acting as a metabolic sensor for AMP levels. There are multiple genes for each subunit and the rat liver AMPK alpha1 and alpha2 catalytic subunits are associated with beta1 and gamma1 noncatalytic subunits. We find that the isolated gamma1 subunit is N-terminally acetylated with no other posttranslational modification. The isolated beta1 subunit is N-terminally myristoylated. Transfection of COS cells with AMPK subunit cDNAs containing a nonmyristoylatable beta1 reduces, but does not eliminate, membrane binding of AMPK heterotrimer. The isolated beta1 subunit is partially phosphorylated at three sites, Ser24/25, Ser182, and Ser108. The Ser24/25 and Ser108 sites are substoichiometrically phosphorylated and can be autophosphorylated in vitro. The Ser-Pro site in the sequence LSSS182PPGP is stoichiometrically phosphorylated, and no additional phosphate is incorporated into this site with autophosphorylation. Based on labeling studies in transfected cells, we conclude that alpha1 Thr172 is a major, although not exclusive, site of both basal and stimulated alpha1 phosphorylation by an upstream AMPK kinase.
Mitchelhill
K I
KI
St. Vincent's Institute of Medical Research, 41 Victoria Parade, Fitzroy, Victoria 3065 Australia.
Michell
B J
BJ
House
C M
CM
Stapleton
D
D
Dyck
J
J
Gamble
J
J
Ullrich
C
C
Witters
L A
LA
Kemp
B E
BE
D055372:AMP-Activated Protein Kinases; D000595:Amino Acid Sequence; D000818:Animals; D002384:Catalysis; D008099:Liver / Q000201:enzymology; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000737:chemistry / Q000378:metabolism*; D019814:Myristic Acid; D009227:Myristic Acids / Q000378:metabolism; D010449:Peptide Mapping; D010766:Phosphorylation; D011494:Protein Kinases / Q000737:chemistry / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D017346:Protein Serine-Threonine Kinases; D051381:Rats; D013347:Subcellular Fractions / Q000201:enzymology
D009097:Multienzyme Complexes; D009227:Myristic Acids; D019814:Myristic Acid; D011494:Protein Kinases; D017346:Protein Serine-Threonine Kinases; D055372:AMP-Activated Protein Kinases