title: Serine carboxypeptidase-like acyltransferases from plants.
abstract: Serine carboxypeptidase-like (SCPL) acyltransferases facilitate transacylation reactions using energy-rich 1-O-β-glucose esters in the synthesis of an array of bioactive compounds and are associated with the diversification of plant natural products. SCPL acyltransferases have evolved from a hydrolytic ancestor by adapting functional elements of the proteases such as the catalytic triad, oxyanion hole, and substrate recognition H-bond network to their new function. As vacuolar proteins, SCPL acyltransferases define an alternative cellular route of transacylation spatially separated from the cytoplasmic enzymes of the BAHD acyltransferase family named according to the first characterized members (BEAT, AHCT, HCBT, and DAT). Recent efforts in cloning and characterization led to the identification of diagnostic peptides for SCPL acyltransferases, enabling the detection of candidate genes in several plant genomes. Detailed biochemical analysis of SCPL acyltransferases is strongly dependent on comprehensive heterologous expression systems, efficient protein purification protocols, and the supply of appropriate substrates. This chapter describes some useful techniques and strategies for identification and characterization of SCPL acyltransferases.
authors.affiliation: Department of Metabolic Biology, John Innes Centre, Norwich Research Park, Norwich, United Kingdom.
authors.forename: Sam T
authors.initials: ST
authors.lastname: Mugford
authors.forename: Carsten
authors.initials: C
authors.lastname: Milkowski
mesh_terms: D000215:Acylation; D000217:Acyltransferases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D017360:Arabidopsis / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D055162:Biocatalysis; D002268:Carboxypeptidases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D019143:Evolution, Molecular; D015870:Gene Expression; D006860:Hydrogen Bonding; D010802:Phylogeny; D010940:Plant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D013379:Substrate Specificity; D014617:Vacuoles / Q000201:enzymology* / Q000235:genetics
chemical_list: D010940:Plant Proteins; D000217:Acyltransferases; D002268:Carboxypeptidases; C022633:serine carboxypeptidase