Of Arginine-Specific Adenosine-5′-Diphosphate-Ribosyltransferase by Capillary Electrophoresis

✍ Scribed by M. Tsuchiya; H. Osago; M. Shimoyama

• Publisher: Elsevier Science
• Year: 1995
• Tongue: English
• Weight: 280 KB
• Volume: 224
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.1995.1076

✦ Synopsis

Using high-voltage capillary electrophoresis we detected ADP-ribosylarginine, a product of ADP-ribosylation reaction catalyzed by arginine-specific ADPribosyltransferase in the presence of NAD and (L)-arginine. The authentic ADP-ribosylarginine, detected by its ultraviolet absorbance at (254 \mathrm{~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 \mathrm{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. 1995 Academic Press, Inc.