title: Assay of NAT Activity.
abstract: In animal tissues, N-acyltransferase (NAT) catalyzes the first reaction in the biosynthetic pathway of bioactive N-acylethanolamines, in which an acyl chain is transferred from the sn-1 position of the donor phospholipid, such as phosphatidylcholine, to the amino group of phosphatidylethanolamine, resulting in the formation of N-acylphosphatidylethanolamine. NAT has long been known to be stimulated by Ca(2+), and hence it has been referred to as Ca(2+)-dependent NAT. On the other hand, members of the phospholipase A/acyltransferase (PLA/AT) family (also known as HRAS-like suppressor family) show Ca(2+)-independent NAT activity. In this chapter, we describe (1) partial purification of Ca(2+)-dependent NAT from rat brain, (2) purification of recombinant PLA/AT-2, and (3) NAT assay using radiolabeled substrate.
authors.affiliation: Department of Biochemistry, Kagawa University School of Medicine, 1750-1 Ikenobe, Miki, Kagawa, 761-0793, Japan.
authors.forename: Toru
authors.initials: T
authors.lastname: Uyama
authors.affiliation: Department of Biochemistry, Kagawa University School of Medicine, 1750-1 Ikenobe, Miki, Kagawa, 761-0793, Japan. nueda@med.kagawa-u.ac.jp.
authors.forename: Natsuo
authors.initials: N
authors.lastname: Ueda
mesh_terms: D000217:Acyltransferases / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D001921:Brain / Q000378:metabolism; D019556:COS Cells; D002118:Calcium / Q000378:metabolism; D002522:Chlorocebus aethiops; D004789:Enzyme Activation; D057075:Enzyme Assays; D010714:Phosphatidylethanolamines; D010741:Phospholipases A / Q000378:metabolism; D051381:Rats
chemical_list: C000609813:N-acylphosphatidylethanolamine; D010714:Phosphatidylethanolamines; D000217:Acyltransferases; D010741:Phospholipases A; D002118:Calcium
keywords: COS-7 cell; Ca-NAT; HRASLS family; N-acylphosphatidylethanolamine; N-acyltransferase; NAPE; PLA/AT family; Radioisotope; Rat brain; Thin-layer chromatography