Acyl-protein thioesterase 2 catalyzes the deacylation of peripheral membrane-associated GAP-43.

An acylation/deacylation cycle is necessary to maintain the steady-state subcellular distribution and biological activity of S-acylated peripheral proteins. Despite the progress that has been made in identifying and characterizing palmitoyltransferases (PATs), much less is known about the thioestera...

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Autores principales:	Vanesa M Tomatis, Alejandra Trenchi, Guillermo A Gomez, Jose L Daniotti
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2010
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/49701025ec23476695801271838181d5
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spelling	oai:doaj.org-article:49701025ec23476695801271838181d52021-11-18T07:02:19ZAcyl-protein thioesterase 2 catalyzes the deacylation of peripheral membrane-associated GAP-43.1932-620310.1371/journal.pone.0015045https://doaj.org/article/49701025ec23476695801271838181d52010-11-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21152083/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203An acylation/deacylation cycle is necessary to maintain the steady-state subcellular distribution and biological activity of S-acylated peripheral proteins. Despite the progress that has been made in identifying and characterizing palmitoyltransferases (PATs), much less is known about the thioesterases involved in protein deacylation. In this work, we investigated the deacylation of growth-associated protein-43 (GAP-43), a dually acylated protein at cysteine residues 3 and 4. Using fluorescent fusion constructs, we measured in vivo the rate of deacylation of GAP-43 and its single acylated mutants in Chinese hamster ovary (CHO)-K1 and human HeLa cells. Biochemical and live cell imaging experiments demonstrated that single acylated mutants were completely deacylated with similar kinetic in both cell types. By RT-PCR we observed that acyl-protein thioesterase 1 (APT-1), the only bona fide thioesterase shown to mediate deacylation in vivo, is expressed in HeLa cells, but not in CHO-K1 cells. However, APT-1 overexpression neither increased the deacylation rate of single acylated GAP-43 nor affected the steady-state subcellular distribution of dually acylated GAP-43 both in CHO-K1 and HeLa cells, indicating that GAP-43 deacylation is not mediated by APT-1. Accordingly, we performed a bioinformatic search to identify putative candidates with acyl-protein thioesterase activity. Among several candidates, we found that APT-2 is expressed both in CHO-K1 and HeLa cells and its overexpression increased the deacylation rate of single acylated GAP-43 and affected the steady-state localization of diacylated GAP-43 and H-Ras. Thus, the results demonstrate that APT-2 is the protein thioesterase involved in the acylation/deacylation cycle operating in GAP-43 subcellular distribution.Vanesa M TomatisAlejandra TrenchiGuillermo A GomezJose L DaniottiPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 5, Iss 11, p e15045 (2010)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Vanesa M Tomatis Alejandra Trenchi Guillermo A Gomez Jose L Daniotti Acyl-protein thioesterase 2 catalyzes the deacylation of peripheral membrane-associated GAP-43.
description	An acylation/deacylation cycle is necessary to maintain the steady-state subcellular distribution and biological activity of S-acylated peripheral proteins. Despite the progress that has been made in identifying and characterizing palmitoyltransferases (PATs), much less is known about the thioesterases involved in protein deacylation. In this work, we investigated the deacylation of growth-associated protein-43 (GAP-43), a dually acylated protein at cysteine residues 3 and 4. Using fluorescent fusion constructs, we measured in vivo the rate of deacylation of GAP-43 and its single acylated mutants in Chinese hamster ovary (CHO)-K1 and human HeLa cells. Biochemical and live cell imaging experiments demonstrated that single acylated mutants were completely deacylated with similar kinetic in both cell types. By RT-PCR we observed that acyl-protein thioesterase 1 (APT-1), the only bona fide thioesterase shown to mediate deacylation in vivo, is expressed in HeLa cells, but not in CHO-K1 cells. However, APT-1 overexpression neither increased the deacylation rate of single acylated GAP-43 nor affected the steady-state subcellular distribution of dually acylated GAP-43 both in CHO-K1 and HeLa cells, indicating that GAP-43 deacylation is not mediated by APT-1. Accordingly, we performed a bioinformatic search to identify putative candidates with acyl-protein thioesterase activity. Among several candidates, we found that APT-2 is expressed both in CHO-K1 and HeLa cells and its overexpression increased the deacylation rate of single acylated GAP-43 and affected the steady-state localization of diacylated GAP-43 and H-Ras. Thus, the results demonstrate that APT-2 is the protein thioesterase involved in the acylation/deacylation cycle operating in GAP-43 subcellular distribution.
format	article
author	Vanesa M Tomatis Alejandra Trenchi Guillermo A Gomez Jose L Daniotti
author_facet	Vanesa M Tomatis Alejandra Trenchi Guillermo A Gomez Jose L Daniotti
author_sort	Vanesa M Tomatis
title	Acyl-protein thioesterase 2 catalyzes the deacylation of peripheral membrane-associated GAP-43.
title_short	Acyl-protein thioesterase 2 catalyzes the deacylation of peripheral membrane-associated GAP-43.
title_full	Acyl-protein thioesterase 2 catalyzes the deacylation of peripheral membrane-associated GAP-43.
title_fullStr	Acyl-protein thioesterase 2 catalyzes the deacylation of peripheral membrane-associated GAP-43.
title_full_unstemmed	Acyl-protein thioesterase 2 catalyzes the deacylation of peripheral membrane-associated GAP-43.
title_sort	acyl-protein thioesterase 2 catalyzes the deacylation of peripheral membrane-associated gap-43.
publisher	Public Library of Science (PLoS)
publishDate	2010
url	https://doaj.org/article/49701025ec23476695801271838181d5
work_keys_str_mv	AT vanesamtomatis acylproteinthioesterase2catalyzesthedeacylationofperipheralmembraneassociatedgap43 AT alejandratrenchi acylproteinthioesterase2catalyzesthedeacylationofperipheralmembraneassociatedgap43 AT guillermoagomez acylproteinthioesterase2catalyzesthedeacylationofperipheralmembraneassociatedgap43 AT joseldaniotti acylproteinthioesterase2catalyzesthedeacylationofperipheralmembraneassociatedgap43
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Acyl-protein thioesterase 2 catalyzes the deacylation of peripheral membrane-associated GAP-43.

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