Molecular basis of fatty acid selectivity in the zDHHC family of S-acyltransferases revealed by click chemistry.
S-acylation is a major posttranslational modification, catalyzed by the zinc finger DHHC domain containing (zDHHC) enzyme family. S-acylated proteins can be modified by different fatty acids; however, very little is known about how zDHHC enzymes contribute to acyl chain heterogeneity. Here, we used fatty acid-azide/alkyne labeling of mammalian cells, showing their transformation into acyl-CoAs and subsequent click chemistry-based detection, to demonstrate that zDHHC enzymes have marked differences in their fatty acid selectivity. This difference in selectivity was apparent even for highly related enzymes, such as zDHHC3 and zDHHC7, which displayed a marked difference in their ability to use C18:0 acyl-CoA as a substrate. Furthermore, we identified isoleucine-182 in transmembrane domain 3 of zDHHC3 as a key determinant in limiting the use of longer chain acyl-CoAs by this enzyme. This study uncovered differences in the fatty acid selectivity profiles of cellular zDHHC enzymes and mapped molecular determinants governing this selectivity.
Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow G4 0RE, United Kingdom.
Jennifer
J
Greaves
WestCHEM, Department of Pure and Applied Chemistry, University of Strathclyde, Glasgow G1 1XL, United Kingdom.
Kevin R
KR
Munro
WestCHEM, Department of Pure and Applied Chemistry, University of Strathclyde, Glasgow G1 1XL, United Kingdom.
Stuart C
SC
Davidson
WestCHEM, Department of Pure and Applied Chemistry, University of Strathclyde, Glasgow G1 1XL, United Kingdom.
Matthieu
M
Riviere
WestCHEM, Department of Pure and Applied Chemistry, University of Strathclyde, Glasgow G1 1XL, United Kingdom.
Justyna
J
Wojno
Biomedical Sciences Research Complex, Schools of Biology and Chemistry, University of St. Andrews, St. Andrews, Fife KY16 9ST, United Kingdom.
Terry K
TK
Smith
WestCHEM, Department of Pure and Applied Chemistry, University of Strathclyde, Glasgow G1 1XL, United Kingdom.
Nicholas C O
NC
Tomkinson
Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow G4 0RE, United Kingdom; Luke.Chamberlain@strath.ac.uk.
Luke H
LH
Chamberlain
D000214:Acyl Coenzyme A / Q000378:metabolism; D000215:Acylation / Q000502:physiology; D000217:Acyltransferases / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D057930:Click Chemistry / Q000379:methods; D005227:Fatty Acids / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D013379:Substrate Specificity / Q000502:physiology; D016335:Zinc Fingers / Q000502:physiology
D000214:Acyl Coenzyme A; D005227:Fatty Acids; D008565:Membrane Proteins; D000217:Acyltransferases
S-acylation; acyl CoA; click chemistry; palmitoylation; zDHHC enzyme