title: Purification and characterization of recombinant protein acyltransferases.
abstract: Palmitoylation enhances membrane association and plays a role in the subcellular trafficking and signaling function of proteins. Unlike other forms of protein lipidation, such as prenylation and myristoylation, palmitoylation is reversible and can therefore play a regulatory role. Enzyme activities have recently been described in mammals and yeast that carry out the palmitoylation of protein substrates. Protein acyltransferases (PATs) transfer a palmitoyl moiety derived from palmitoyl-CoA to a free thiol of a substrate protein to create a labile thioester linkage. Biochemical characterization and kinetic analysis of this new family of enzymes requires methods to purify PATs and their substrates, as well as methods to assay PAT activity. We describe a series of methods using yeast and bacterial expression systems to study protein acyltransferases.
authors.affiliation: Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI 53226, USA.
authors.forename: Cheryl
authors.initials: C
authors.lastname: Budde
authors.forename: Marissa J
authors.initials: MJ
authors.lastname: Schoenfish
authors.forename: Maurine E
authors.initials: ME
authors.lastname: Linder
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Deschenes
mesh_terms: D000217:Acyltransferases / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification*; D019308:Palmitic Acid / Q000378:metabolism; D011994:Recombinant Proteins / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae
chemical_list: D011994:Recombinant Proteins; D019308:Palmitic Acid; D000217:Acyltransferases