2-bromopalmitate reduces protein deacylation by inhibition of acyl-protein thioesterase enzymatic activities

Pedro, Maria P., Vilcaes, Aldo A., Tomatis, Vanesa M., Oliveira, Rafael G., Gomez, Guillermo A. and Daniotti, Jose L. (2013) 2-bromopalmitate reduces protein deacylation by inhibition of acyl-protein thioesterase enzymatic activities. PLoS One, 8 10: e75232.1-e75232.11. doi:10.1371/journal.pone.0075232


Author Pedro, Maria P.
Vilcaes, Aldo A.
Tomatis, Vanesa M.
Oliveira, Rafael G.
Gomez, Guillermo A.
Daniotti, Jose L.
Title 2-bromopalmitate reduces protein deacylation by inhibition of acyl-protein thioesterase enzymatic activities
Journal name PLoS One   Check publisher's open access policy
ISSN 1932-6203
Publication date 2013-10-01
Sub-type Article (original research)
DOI 10.1371/journal.pone.0075232
Open Access Status DOI
Volume 8
Issue 10
Start page e75232.1
End page e75232.11
Total pages 11
Place of publication San Francisco, CA, United States
Publisher Public Library of Science
Language eng
Abstract S-acylation, the covalent attachment of palmitate and other fatty acids on cysteine residues, is a reversible post-translational modification that exerts diverse effects on protein functions. S-acylation is catalyzed by protein acyltransferases (PAT), while deacylation requires acyl-protein thioesterases (APT), with numerous inhibitors for these enzymes having already been developed and characterized. Among these inhibitors, the palmitate analog 2-brompalmitate (2-BP) is the most commonly used to inhibit palmitoylation in cells. Nevertheless, previous results from our laboratory have suggested that 2-BP could affect protein deacylation. Here, we further investigated in vivo and in vitro the effect of 2-BP on the acylation/deacylation protein machinery, with it being observed that 2-BP, in addition to inhibiting PAT activity in vivo, also perturbed the acylation cycle of GAP-43 at the level of depalmitoylation and consequently affected its kinetics of membrane association. Furthermore, 2-BP was able to inhibit in vitro the enzymatic activities of human APT1 and APT2, the only two thioesterases shown to mediate protein deacylation, through an uncompetitive mechanism of action. In fact, APT1 and APT2 hydrolyzed both the monomeric form as well as the micellar state of the substrate palmitoyl-CoA. On the basis of the obtained results, as APTs can mediate deacylation on membrane bound and unbound substrates, this suggests that the access of APTs to the membrane interface is not a necessary requisite for deacylation. Moreover, as the enzymatic activity of APTs was inhibited by 2-BP treatment, then the kinetics analysis of protein acylation using 2-BP should be carefully interpreted, as this drug also inhibits protein deacylation.
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Queensland Brain Institute Publications
Official 2014 Collection
Institute for Molecular Bioscience - Publications
 
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