Identification of Acyl Protein Thioesterases 1 and 2 as the Cellular Targets of the Ras‐Signaling Modulators Palmostatin B and M

Rusch, Marion; Zimmermann, T.; Bürger, Marco; Görmer, Kristina; Triola, Gemma; Brockmeyer, Andreas; Janning, Petra; Böttcher, Thomas; Sieber, Stephan A.; Vetter, Ingrid R.; Hedberg, Christian; Waldmann, Herbert

doi:10.1002/anie.201102967

Cited by 106 publications

(66 citation statements)

References 26 publications

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“…Palmostatin M inhibits the eukaryotic hydrolase enzymes APT1 and APT2 by blocking their Ras GTPase depalmitoylation activity and in turn altering Ras GTPase localization and activity [26,27]. The β-lactone Palmostatin M may also have off-target effects on bacterial serine hydrolases with a similar active site.…”

Section: Resultsmentioning

confidence: 99%

Exploring Anti-Bacterial Compounds against Intracellular Legionella

et al. 2013

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Legionella pneumophila is a ubiquitous fresh-water bacterium which reproduces within its erstwhile predators, environmental amoeba, by subverting the normal pathway of phagocytosis and degradation. The molecular mechanisms which confer resistance to amoeba are apparently conserved and also allow replication within macrophages. Thus, L. pneumophila can act as an ‘accidental’ human pathogen and cause a severe pneumonia known as Legionnaires’ disease. The intracellular localisation of L. pneumophila protects it from some antibiotics, and this fact must be taken into account to develop new anti-bacterial compounds. In addition, the intracellular lifestyle of L. pneumophila may render the bacteria susceptible to compounds diminishing bacterial virulence and decreasing intracellular survival and replication of this pathogen. The development of a single infection cycle intracellular replication assay using GFP-producing L. pneumophila and Acanthamoeba castellanii amoeba is reported here. This fluorescence-based assay allows for continuous monitoring of intracellular replication rates, revealing the effect of bacterial gene deletions or drug treatment. To examine how perturbations of the host cell affect L. pneumophila replication, several known host-targeting compounds were tested, including modulators of cytoskeletal dynamics, vesicle scission and Ras GTPase localisation. Our results reveal a hitherto unrealized potential antibiotic property of the β-lactone-based Ras depalmitoylation inhibitor palmostatin M, but not the closely related inhibitor palmostatin B. Further characterisation indicated that this compound caused specific growth inhibition of Legionella and Mycobacterium species, suggesting that it may act on a common bacterial target.

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Section: Resultsmentioning

confidence: 99%

Exploring Anti-Bacterial Compounds against Intracellular Legionella

et al. 2013

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“…A PT2 has also been reported to depalmitoylate H-Ras like APT1. However, it has more efficient catalytic activity than APT1 in depalmitoylating a biologically active semisynthetic N-Ras in vitro [105]. Overexpression of APT2, but not APT1, causes rapid depalmitoylation of the GAP-43 [106], demonstrating the substrate specificity of APT2.…”

Section: Acyl Protein Thioesterase 1 (Apts)mentioning

confidence: 99%

Palmitoylation in Alzheimer⿿s disease and other neurodegenerative diseases

Cho

Park

2016

Pharmacological Research

115

108

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Posttranslational modifications of proteins are important regulatory processes endowing the proteins functional complexity. Over the last decade, numerous studies have shed light on the roles of palmitoylation, one of the most common lipid modifications, in various aspects of neuronal functions. Major players regulating palmitoylation are the enzymes that mediate palmitoylation and depalmitoylation which are palmitoyl acyltransferases (PATs) and protein thioesterases, respectively. In this review, we will provide and discuss current understandings on palmitoyation/depalmitoylation control mediated by PATs and/or protein thioesterases for neuronal functions in general and also for Alzheimer's disease in particular, and other neurodegenerative diseases such as Huntington's disease, schizophrenia and intellectual disability.

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“…Interestingly, siRNA knockdown of APT1 did not statistically alter the ratio of golgi to plasma membrane NRas. This suggests either insufficient APT1 knockdown, or that Palmostatin B inhibits other protein thioesterases, such as APT2 ( K i = 34 nM) 18 that may contribute to NRas processing. Further experiments showed that addition of 30 - 100 µM of Palmostatin B dose-dependently impedes the growth of HRas or NRas transduced hematopoietic cells, but does not affect cells transduced with a non-palmitoylated isoform of KRas 29 .…”

Section: β-Lactonesmentioning

confidence: 99%

“…Palmostatin M similarly induces NRas mis-localization from the plasma membrane and partially restores E-Cadherin localization at cell-cell junctions in HRas-transformed MDCK cells 38 . Because β-lactones form a semi-stable covalent complex with their targets, an alkynyl Palmostatin M analogue was synthesized to annotate potential cellular targets 18 . Lysates were labeled with the alkynyl probes, conjugated to a tri-functional rhodamine/biotin-azide by copper-catalyzed click chemistry, enriched with streptavidin, and analyzed by in-gel fluorescence or mass spectrometry to identify candidate target proteins.…”

Section: β-Lactonesmentioning

confidence: 99%

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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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Identification of Acyl Protein Thioesterases 1 and 2 as the Cellular Targets of the Ras‐Signaling Modulators Palmostatin B and M

Abstract: Finding the target: activity-based proteomic profiling probes based on the depalmitoylation inhibitors palmostatin B and M have been synthesized and were found to target acyl protein thioesterase 1 (APT1) and 2 (APT2) in cells.

Cited by 106 publications

References 26 publications

Exploring Anti-Bacterial Compounds against Intracellular Legionella

Exploring Anti-Bacterial Compounds against Intracellular Legionella

Palmitoylation in Alzheimer⿿s disease and other neurodegenerative diseases

Acyl protein thioesterase inhibitors as probes of dynamicS-palmitoylation

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