Radiometric Assay of ABHD2 Activity

Methods Mol Biol. 2023:2576:299-305. doi: 10.1007/978-1-0716-2728-0_25.

Abstract

The α,β-hydrolase fold-containing protein 2 (ABHD2) is a serine hydrolase, responsible for the cleavage of endogenous 2-arachidonoylglycerol (2-AG). ABHD2 is activated by progesterone, thus, it is considered a nonnuclear receptor of this steroid hormone that terminates its biological effects. The products of ABHD2-catalyzed cleavage by the natural substrate 2-AG are glycerol and arachidonic acid; here, instead of 2-AG, the radioactive substrate 2-oleoyl-[3H]glycerol has been used as already done in various acylglycerol lipase activity assays. The amount of [3H]glycerol released allows to measure ABHD2 enzymatic activity.

Keywords: ABHD2; Acylglycerol lipase; Bioactive lipids; Endocannabinoids; Progesterone receptor; U2OS cells.

MeSH terms

  • Arachidonic Acid
  • Arachidonic Acids* / metabolism
  • Endocannabinoids
  • Glycerides* / metabolism
  • Glycerol
  • Lipase / metabolism
  • Progesterone
  • Serine

Substances

  • Arachidonic Acids
  • Endocannabinoids
  • Glycerides
  • Arachidonic Acid
  • Serine
  • Progesterone
  • Lipase
  • Glycerol