ADP-ribosylation suppresses phosphorylation of the L-type pyruvate kinase.
L-type pyruvate kinase (EC 2.7.1.40) purified from pig liver was ADP-ribosylated by incubation with NAD and ADP-ribosyltransferase purified from hen liver nuclei. Maximal incorporation of the ADP-ribose moiety from NAD into the L-type pyruvate kinase was 0.98 mol/mol of subunit. The Km values for NAD and L-type pyruvate kinase were 0.17 mM and 9.7 microM, respectively. ADP-ribosylation of the L-type pyruvate kinase resulted in suppression of the subsequent phosphorylation catalyzed by cAMP-dependent protein kinase. The ADP-ribosylation-induced suppression of phosphorylation of the L-type pyruvate kinase also resulted in suppression of the phosphorylation-induced inactivation. Amino acid analysis, after exhaustive sequential digestion of ADP-ribosyl-L-type pyruvate kinase with pepsin, aminopeptidase M and carboxy-peptidase B showed arginine to be the ADP-ribose-accepting amino acid. These results together with finding of the ADP-ribosyltransferase activity in mammalian liver cytosol (Moss, J. and Stanley, S.J. (1981) J. Biol. Chem. 256, 7830-7833) suggest that ADP-ribosylation may participate in the regulation of the L-type pyruvate kinase activity through changes in the rate of phosphorylation.
Matsuura
R
R
Department of Biochemistry, Shimane Medical University, Izumo, Japan.
Tanigawa
Y
Y
Tsuchiya
M
M
Mishima
K
K
Yoshimura
Y
Y
Shimoyama
M
M
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism*; D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives; D002645:Chickens; D066298:In Vitro Techniques; D008099:Liver / Q000201:enzymology; D010430:Pentosyltransferases / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism; D011770:Pyruvate Kinase / Q000378:metabolism*
D010750:Phosphoproteins; D000246:Adenosine Diphosphate Ribose; D001120:Arginine; D010430:Pentosyltransferases; D011494:Protein Kinases; D011770:Pyruvate Kinase