Assay of arginine-specific adenosine-5'-diphosphate-ribosyltransferase by capillary electrophoresis.
Using high-voltage capillary electrophoresis we detected ADP-ribosylarginine, a product of ADP-ribosylation reaction catalyzed by arginine-specific ADP-ribosyltransferase in the presence of NAD and L-arginine. The authentic ADP-ribosylarginine, detected by its ultraviolet absorbance at 254 nm, had a different retention time from NAD or nicotinamide. When the ADP-ribosylation reaction products were analyzed, the peak corresponding to ADP-ribosylarginine increased with incubation time and in an enzyme-dose-dependent manner. The lower limit of detection was 0.3 pmol, a value 100 times lower than that obtained with the reversed-phase high-performance liquid chromatography assay described previously. Using the capillary electrophoresis system, a thiol-independent ADP-ribosyltransferase activity was detected in chicken spleen cell membrane. Since the capillary electrophoresis assay for ADP-ribosylarginine is simpler, faster, and less expensive than the high-performance liquid chromatography assay, determination of arginine-specific ADP-ribosyltransferase activity in animal tissues will be facilitated.
Tsuchiya
M
M
Department of Biochemistry, Shimane Medical University, Japan.
Osago
H
H
Shimoyama
M
M
D036002:ADP Ribose Transferases / Q000032:analysis* / Q000378:metabolism; D000246:Adenosine Diphosphate Ribose / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism; D000818:Animals; D001120:Arginine / Q000378:metabolism; D002645:Chickens; D004586:Electrophoresis / Q000379:methods; D009249:NADP / Q000378:metabolism
D000246:Adenosine Diphosphate Ribose; D009249:NADP; D001120:Arginine; C043357:ADP-ribosylarginine; D036002:ADP Ribose Transferases