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Identification of deubiquitinases controlled by extracellular matrix stiffness in cancer cells

ABSTRACT: The project was aimed at identifying deubiquitinases (DUBs) involved in cancer cell mechanosignaling. Lysates from cancer cells grown on collagen matrices with varying stiffnesses were incubated with an activity-based HA-tagged ubiquitin probe that covalently binds DUB active site. To identify DUBs differentially activated on soft and stiff collagen hydrogels, lysates were immunoprecipitated with anti-HA agarose beads. Immunoprecipitates were further identified by LC-MS/MS analysis.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Melanocyte

DISEASE(S): Skin Cancer

SUBMITTER: AUDEBERT Stephane

LAB HEAD: DECKERT Marcel, PhD

PROVIDER: PXD054010 | Pride | 2025-08-22

REPOSITORIES: Pride

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Project description:Mycobacteria are known for their unique and complex cell wall architecture, characterized by distinct lipid-rich outer layers, making lipid metabolism a central point of their physiological processes. One crucial aspect of all mycobacteria is their ability to metabolize host lipids and to accumulate the resulting neutral lipids in their own cytoplasm in the form of intrabacterial lipid inclusions (ILI). These lipid-rich organelles consist of a hydrophobic core containing neutral lipids, essentially triacylglycerol (TAG), surrounded by a phospholipid monolayer, associated with numerous proteins. These ILI not only serve as reservoirs for energy storage and provide a platform for the synthesis and maintenance of lipids, which are essential for the adaptability and survival of mycobacteria under various environmental stresses. How these mycobacteria acquire host lipids and how ILI are formed remain important and largely unanswered questions. More generally, the anabolism and catabolism of ILI enhance the survival and long-term persistence of mycobacteria and thus represent two major processes controlled by a number of enzymes and regulatory proteins that play an essential role in the biosynthesis, modification, and utilization of these lipid inclusions. In this context, we report here the synthesis of Oxadiazolone (OX) and the Cyclophostin & Cyclipostins (CyC) activity-based probes, and their use for the direct capture of target proteins in M. abscessus growing under carbon excess and nitrogen-deprived in vitro conditions that promote TAG production and ILI-positive phenotype, via bio-orthogonal click-chemistry activity-based protein profiling (CC-ABPP). This approach led to the identification of a set of 65 enzymes potentially involved in the global processes related to ILI anabolism. Among these enzymes, the long-chain-fatty-acid--CoA ligase MAB_1978c/FadD15 has been validated not only as a pivotal enzyme colocalized on ILI, but above all as a major contributor in ILI formation in M. abscessus.

			Action	DRS
	Results.zip	Other
	Soft_Rep1.raw	Raw
	Soft_Rep1_20211112155256.raw	Raw
	Soft_Rep1_20211112183213.raw	Raw
	Soft_Rep2.raw	Raw

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Identification of deubiquitinases controlled by extracellular matrix stiffness in cancer cells

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