Phosphorylation of rat muscle acetyl-CoA carboxylase by AMP-activated protein kinase and protein kinase A.
This study was designed to compare functional effects of phosphorylation of muscle acetyl-CoA carboxylase (ACC) by adenosine 3',5'-cyclic monophosphate-dependent protein kinase (PKA) and by AMP-activated protein kinase (AMPK). Muscle ACC (272 kDa) was phosphorylated and then subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by autoradiography. Functional effects of phosphorylation were determined by measuring ACC activity at different concentrations of each of the substrates and of citrate, an activator of the enzyme. The maximal velocity (Vmax) and the Michaelis constants (Km) for ATP, acetyl-CoA, and bicarbonate were unaffected by phosphorylation by PKA. Phosphorylation by AMPK increased the Km for ATP and acetyl-CoA. Sequential phosphorylation by PKA and AMPK, first without label and second with label, appeared to reduce the extent of label incorporation, regardless of the order. The activation constant (Ka) for citrate activation was increased to the same extent by AMPK phosphorylation, regardless of previous or subsequent phosphorylation by PKA. Thus muscle ACC can be phosphorylated by PKA but with no apparent functional effects on the enzyme. AMPK appears to be the more important regulator of muscle ACC.
Winder
W W
WW
Department of Zoology, Brigham Young University, Provo, Utah 84602, USA. winderw@acd1.byu.edu
Wilson
H A
HA
Hardie
D G
DG
Rasmussen
B B
BB
Hutber
C A
CA
Call
G B
GB
Clayton
R D
RD
Conley
L M
LM
Yoon
S
S
Zhou
B
B
D000103:Acetyl-CoA Carboxylase / Q000187:drug effects* / Q000378:metabolism; D000249:Adenosine Monophosphate / Q000494:pharmacology*; D000818:Animals; D017868:Cyclic AMP-Dependent Protein Kinases / Q000187:drug effects* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D018482:Muscle, Skeletal / Q000187:drug effects* / Q000378:metabolism; D010766:Phosphorylation; D011494:Protein Kinases / Q000187:drug effects* / Q000378:metabolism; D051381:Rats
D000249:Adenosine Monophosphate; D011494:Protein Kinases; D017868:Cyclic AMP-Dependent Protein Kinases; D000103:Acetyl-CoA Carboxylase