A Highly Sensitive High-Performance Liquid Chromatography– Fluorometric Method for the Assay of Peptidylarginine Deiminase Activity

✍ Scribed by Toshiyuki Chikuma; Michiyuki Yamada; Ayumi Tsuda; Masaru Yamamoto; Katsuhiko Nakashima; Ryuichi Yajima; Takeshi Kato

• Publisher: Elsevier Science
• Year: 2000
• Tongue: English
• Weight: 68 KB
• Volume: 285
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.2000.4769

✦ Synopsis

The activity of peptidylarginine deiminase (PAD) has generally been assayed by a colorimetric method using N-benzoyl-L-arginine ethyl ester (BAEE) and Nbenzoyl-L-arginine (Bz-L-Arg) as the substrates. The widespread occurrence of citrulline and urea in tissues makes use of this method difficult, especially for small samples. We developed a highly sensitive highperformance liquid chromatography method with Ndansyl-glycyl-L-arginine as the substrate. This method was sensitive enough to determine previously undetectable activity of PAD in HL-60 cells. Two types of PAD (HL-60 cell and brain PAD) could be distinguished by differential competition, using either BAEE or Bz-L-Arg as a preferential substrate in the assay. These data indicate that the present method is applicable to many tissues.