title: The Golgi S-acylation machinery comprises zDHHC enzymes with major differences in substrate affinity and S-acylation activity.
abstract: S-acylation, the attachment of fatty acids onto cysteine residues, regulates protein trafficking and function and is mediated by a family of zDHHC enzymes. The S-acylation of peripheral membrane proteins has been proposed to occur at the Golgi, catalyzed by an S-acylation machinery that displays little substrate specificity. To advance understanding of how S-acylation of peripheral membrane proteins is handled by Golgi zDHHC enzymes, we investigated interactions between a subset of four Golgi zDHHC enzymes and two S-acylated proteins-synaptosomal-associated protein 25 (SNAP25) and cysteine-string protein (CSP). Our results uncover major differences in substrate recognition and S-acylation by these zDHHC enzymes. The ankyrin-repeat domains of zDHHC17 and zDHHC13 mediated strong and selective interactions with SNAP25/CSP, whereas binding of zDHHC3 and zDHHC7 to these proteins was barely detectable. Despite this, zDHHC3/zDHHC7 could S-acylate SNAP25/CSP more efficiently than zDHHC17, whereas zDHHC13 lacked S-acylation activity toward these proteins. Overall the results of this study support a model in which dynamic intracellular localization of peripheral membrane proteins is achieved by highly selective recruitment by a subset of zDHHC enzymes at the Golgi, combined with highly efficient S-acylation by other Golgi zDHHC enzymes.
authors.affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow G4 0RE, United Kingdom.
authors.forename: Kimon
authors.initials: K
authors.lastname: Lemonidis
authors.affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow G4 0RE, United Kingdom.
authors.forename: Oforiwa A
authors.initials: OA
authors.lastname: Gorleku
authors.affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow G4 0RE, United Kingdom.
authors.forename: Maria C
authors.initials: MC
authors.lastname: Sanchez-Perez
authors.affiliation: ZMBP Developmental Genetics, D-72076 Tuebingen, Germany.
authors.forename: Christopher
authors.initials: C
authors.lastname: Grefen
authors.affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow G4 0RE, United Kingdom Luke.Chamberlain@strath.ac.uk.
authors.forename: Luke H
authors.initials: LH
authors.lastname: Chamberlain
mesh_terms: D000123:Acetyltransferases; D000215:Acylation; D000217:Acyltransferases / Q000235:genetics / Q000378:metabolism*; D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism; D001667:Binding, Competitive; D003545:Cysteine / Q000378:metabolism*; D005227:Fatty Acids / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism*; D057809:HEK293 Cells; D050956:HSP40 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D018613:Microscopy, Confocal; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D013379:Substrate Specificity; D050825:Synaptosomal-Associated Protein 25 / Q000235:genetics / Q000378:metabolism
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D005227:Fatty Acids; D050956:HSP40 Heat-Shock Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D050825:Synaptosomal-Associated Protein 25; C088460:cysteine string protein; D000217:Acyltransferases; C574550:ZDHHC3 protein, human; D000123:Acetyltransferases; C000598869:ZDHHC13 protein, human; C473038:ZDHHC17 protein, human; C447819:ZDHHC7 protein, human; D003545:Cysteine