Abstract
Ubiquitin carboxy-terminal hydrolase L1 (UCHL1) is a deubiquitylating enzyme that is proposed as a potential therapeutic target in neurodegeneration, cancer, and liver and lung fibrosis. Herein we report the discovery of the most potent and selective UCHL1 probe (IMP-1710) to date based on a covalent inhibitor scaffold and apply this probe to identify and quantify target proteins in intact human cells. IMP-1710 stereoselectively labels the catalytic cysteine of UCHL1 at low nanomolar concentration in cells. We further demonstrate that potent and selective UCHL1 inhibitors block pro-fibrotic responses in a cellular model of idiopathic pulmonary fibrosis, supporting the potential of UCHL1 as a potential therapeutic target in fibrotic diseases.
Highlights
Ubiquitin carboxy-terminal hydrolase L1 (UCHL1) is a deubiquitylating enzyme that is proposed as a potential therapeutic target in neurodegeneration, cancer, and liver and lung fibrosis
Its biological functions are not yet fully understood, UCHL1 is abundantly expressed in the brain, where it is involved in apoptosis regulation, learning, and memory, while UCHL1 dysregulation is linked to diseases including neurodegeneration,[5] cancers,[6−8] and fibrosis.[9]
Cellular UCHL1 activity was demonstrated in breast cancer cells (Cal51) stably expressing FLAG-UCHL1 using a Ub-vinyl methyl ester probe (HA-UbVME) by a homogeneous time-resolved fluorescence (HTRF) assay.[15]
Summary
Ubiquitin carboxy-terminal hydrolase L1 (UCHL1) is a deubiquitylating enzyme that is proposed as a potential therapeutic target in neurodegeneration, cancer, and liver and lung fibrosis. Cellular potencies were in line with biochemical data (Figure S1), and compound 3 did not inhibit UCHL1 in cells.
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