Work

Chemoproteomic profiling by bioorthogonal probes to reveal the novel targets of acrylamide in microglia

Year: 2024

Type: article

Abstract: Acrylamide (AA) exposure can lead to a variety of adverse health outcomes, with neurotoxicity being the most prevalent. AA is an electrophilic reagent containing α,β-unsaturated amide groups that tend to covalently modify cysteine residues of proteins, but the methods for direct analysis of AA targets in living cells by cysteine profiling are still lacking. Here, we developed a specific bioorthogonal probe, acrylamide-alkyne, for chemical proteomic analysis of AA-binding sites. 754 protein targets of acrylamide were identified in neuromicroglia, of which 96 proteins both highly sensitive and heavily modified were screened. Two new targets, PSMD9 and NDUFA5, which are related to the ubiquitin-proteasome and mitochondrial functional system, respectively, were enriched by the proposed protocol. Site-specific mutation verified that AA could inhibit activity of PSMD9 by modifying C59 of PSMD9 to induce dysfunction of the ubiquitin-proteasome system, and inhibit activity of mitochondrial complex I by modifying C17 of NDUFA5 to trigger apoptosis. Moreover, the study verified the crosstalk between acrylation of PSMD9 and NDUFA5 may induce the mitochondrial activation of neuronal apoptotic mechanisms. Our findings not only provide a biological orthogonal probe tool for studying acrylamide-covalently bound target proteins, but also provide a new idea for investigating molecular mechanism of acrylamide-induced neurotoxicity.

Source: Journal of Hazardous Materials

DOI: 10.1016/j.jhazmat.2024.136760