Purification and characterization of arginine:mono-ADP-ribosylhydrolase from Euglena gracilis Z.
Arginine:mono-ADP-ribosylhydrolase was purified from a protozoan, Euglena gracilis Z, using [32P]mono-ADP-ribosylated actin as a substrate. The enzyme showed molecular mass of 33 kDa both in SDS PAGE and gel filtration, indicating it to be a monomeric protein. It was strongly inhibited by ADP and ADP-ribose and activated by Mg2+, DTT, and 2-mercaptoethanol. These results suggest that it recognizes the ADP-ribose moiety of the modified protein. Since the enzyme activity increased in S phase and late G0 phase in a synchronous dividing culture, the enzyme may function in the regulation of the cell cycle.
Takenaka
S
S
Department of Applied Biological Chemistry, Osaka Prefecture University.
Masuda
W
W
Tsuyama
S
S
Tamura
Y
Y
Miyatake
K
K
Nakano
Y
Y
D000199:Actins / Q000737:chemistry; D000244:Adenosine Diphosphate / Q000737:chemistry; D000818:Animals; D002455:Cell Division; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D005056:Euglena gracilis / Q000201:enzymology*; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008274:Magnesium / Q000737:chemistry; D008623:Mercaptoethanol / Q000737:chemistry; D009699:N-Glycosyl Hydrolases; D015800:Protozoan Proteins / Q000737:chemistry / Q000302:isolation & purification*
D000199:Actins; D015800:Protozoan Proteins; D008623:Mercaptoethanol; D000244:Adenosine Diphosphate; D006026:Glycoside Hydrolases; D009699:N-Glycosyl Hydrolases; C046422:ADP-ribosylarginine hydrolase; D008274:Magnesium