Development and Biological Evaluation of Acyl Protein Thioesterase 1 (APT1) Inhibitors

Deck, Patrick; Pendzialek, Dirk; Biel, Markus; Wagner, Mélanie; Popkirova, Boriana; Ludolph, Björn; Kragol, Goran; Kuhlmann, Jürgen; Giannis, Athanassios; Waldmann, Herbert

doi:10.1002/anie.200462625

Cited by 23 publications

(15 citation statements)

References 20 publications

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“…However, at steady-state conditions 2-BP treatment did not modify the acylation state and membrane binding properties of GAP-43 [34], strongly suggesting that membrane-associated GAP-43 is not being deacylated. However, it should be also considered that deacylated GAP-43 could be a preferred substrate for proteasome degradation [43], [44], avoiding cytosolic accumulation, and/or that 2-BP not only inhibits PATs but also APT activity similarly to that observed using APT-1 inhibitors [45]. To assess these alternatives, cells expressing GAP-43 at steady-state conditions were treated with 2-BP and cycloheximide (CHX) in the presence (+Inh) or absence (-Inh) of proteasomal and lysosomal inhibitors during 0, 3 and 6 h (Fig.…”

Section: Resultsmentioning

confidence: 99%

Acyl-Protein Thioesterase 2 Catalizes the Deacylation of Peripheral Membrane-Associated GAP-43

et al. 2010

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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.

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Section: Resultsmentioning

confidence: 99%

Acyl-Protein Thioesterase 2 Catalizes the Deacylation of Peripheral Membrane-Associated GAP-43

et al. 2010

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“…(5) This finally results in the dissociation of the complex from the membrane. Schematic representation of methods used to introduce natural and modified lipidated C termini on Ras GTPases, and a collection of semisynthetic neo-Ras proteins synthesized via these methods 58,62,64,67,76 FTI and GGTI compounds. Three FTI compounds used in clinical trials are shown.…”

Section: Discussionmentioning

confidence: 99%

“…Until recently no Erf2 orthologs or other proteins featuring Ras S-palmitoylation activity had been found in the genomes of eukaryotes, including the human genome. However, two proteins or protein complexes (DHHC9 and GCP16) that may be involved in Ras palmitoylation have recently been discovered 61,62 .…”

Section: Palmitoylation Of Prenylated Proteinsmentioning

confidence: 99%

“…The relationship between Ras proteins and APT1 has therefore been further studied using a chemical biological approach 64 . In this investigation several semisynthetic Ras proteins, together with lipopeptides and newly developed APT1 inhibitors, were synthesized via combined chemical and biological methods and used as tools, without which the study would have been difficult 62 . These APT1 inhibitors prevented differentiation of PC12 cells by oncogenic N-Ras when N-Ras had to be finally processed within the cell (that is, when N-Ras lacked a palmitoyl anchor).…”

Section: Palmitoylation Of Prenylated Proteinsmentioning

confidence: 99%

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Therapeutic intervention based on protein prenylation and associated modifications

et al. 2006

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In eukaryotic cells, a specific set of proteins are modified by C-terminal attachment of 15-carbon farnesyl groups or 20-carbon geranylgeranyl groups that function both as anchors for fixing proteins to membranes and as molecular handles for facilitating binding of these lipidated proteins to other proteins. Additional modification of these prenylated proteins includes C-terminal proteolysis and methylation, and attachment of a 16-carbon palmitoyl group; these modifications augment membrane anchoring and alter the dynamics of movement of proteins between different cellular membrane compartments. The enzymes in the protein prenylation pathway have been isolated and characterized. Blocking protein prenylation is proving to be therapeutically useful for the treatment of certain cancers, infection by protozoan parasites and the rare genetic disease Hutchinson-Gilford progeria syndrome.Isoprenoids are lipids made of five-carbon blocks, and they constitute one of the main classes of natural products. A subset of isoprenoids called prenyl groups are found on a variety of biological substances, including proteins. Prenylated proteins and peptides are found in most (if not all) eukaryotes. They arise from the post-translational attachment of 15-carbon farnesyl or 20-carbon geranylgeranyl groups to the C-terminal segment of proteins. Protein prenyl groups not only are hydrophobic elements that bind proteins to membranes, but, at least in some cases, they also function as molecular handles that bind to hydrophobic grooves on the surface of soluble protein factors; these factors then remove the prenylated protein from the membrane in a regulated manner. NIH Public Access Author ManuscriptNat Chem Biol. Author manuscript; available in PMC 2010 June 28. Published in final edited form as:Nat Chem Biol. 2006 October ; 2(10): 518-528. doi:10.1038/nchembio818. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author ManuscriptProtein prenylation has been vigorously studied over the past ~15 years because it is found on several signaling proteins (including heterotrimeric G proteins) that connect cell surface receptors to intracellular effectors, and also on Ras proteins, one of the most common oncoproteins found in human tumors. Ras proteins serve as molecular switches at cellular membranes to control propagation of growth signals from cell surface receptors to nuclear transcription factors. Because Ras mutations are important in many human cancers, there has been extensive focus on Ras prenylation. Discovery of protein prenyl groups and structural varietiesThe first reports of prenylated proteins and peptides described the secreted pheromone peptides from jelly fungi 1,2 , whose structure resembles that of the well-known a-factor mating pheromone from baker's yeast (Saccharomyces cerevisiae), which contains a cysteine methylester farnesylated at the C terminus 3 . In the 1980s, studies on the timing of cholesterol biosynthesis with respect to the cell cycle in human cells led to the discovery that a compound deriv...

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“…A peptide-imitating benzodiazepinedione core was fused to S-farnesyl cysteine methyl ester and an acyl-sulfonamide to mimic the serine hydrolase tetrahedral intermediate 34 , yielding a low nanomolar APT1 inhibitor (Compound 10, IC 50 = 27 nM). Intermediates lacking the farnesyl moiety were 5-times less potent (Compound 6, IC 50 = 148 nM), but presumably more soluble, and were tested in cells.…”

Section: Benzodiazepinedionesmentioning

confidence: 99%

Acyl protein thioesterase inhibitors as probes of dynamicS-palmitoylation

Davda

Martin

2014

Med. Chem. Commun.

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Protein palmitoylation describes the hydrophobic post-translational modification of cysteine residues in certain proteins, and is required for the spatial organization and composition of cellular membrane environments. Certain palmitoylated proteins are processed by acyl protein thioesterase (APT) enzymes, which catalyze thioester hydrolysis of palmitoylated cysteine residues. Inhibiting APT enzymes disrupts Ras trafficking and attenuates oncogenic growth signaling, highlighting these enzymes as potential therapeutic targets. As members of the serine hydrolase enzyme family, APT enzymes can be assayed by fluorophosphonate activity-based protein profiling (ABPP) methods, allowing rapid profiling of inhibitor selectivity and potency. In this review, we discuss recent progress in the development of potent and selective inhibitors to APT enzymes, including both competitive and non-competitive chemotypes. These examples highlight how ABPP methods integrate with medicinal chemistry for the discovery and optimization of inhibitors in complex proteomes.

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Development and Biological Evaluation of Acyl Protein Thioesterase 1 (APT1) Inhibitors

Cited by 23 publications

References 20 publications

Acyl-Protein Thioesterase 2 Catalizes the Deacylation of Peripheral Membrane-Associated GAP-43

Acyl-Protein Thioesterase 2 Catalizes the Deacylation of Peripheral Membrane-Associated GAP-43

Therapeutic intervention based on protein prenylation and associated modifications

Acyl protein thioesterase inhibitors as probes of dynamicS-palmitoylation

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