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Querying the In Vitro Proteome Cysteine Reactivity of 8:2 Fluorotelomer Acrylate

ABSTRACT: Despite the phase out of legacy per- and polyfluoroalkyl substances (PFAS), fluorotelomer polymers (FTP) have been used for many applications, notably textile surface coatings. FTPs are of health concerns due to their breakdown into legacy PFAS and the co-occurrence of fluorotelomer acrylate (FTAC) monomers, of which the latter may potentially react with cellular thiols. To investigate these reactions, we employed fluorous-solid phase extraction (FSPE), to enrich peptides covalently modified by 8:2 fluorotelomer acrylate (8:2 FTAC), and coupled it to a modified nano-liquid chromatography method for the identification of in vitro protein adducts using shotgun proteomics. Using this method, over one-hundred unique peptides were detected with 8:2 FTAC modifications, although none of the modified cysteine residues were annotated active site nucleophiles. In parallel, a synthetic C6F13-iodoacetamide (F13-IAM) chemical probe was used to gauge the upper-bound of PFAS-thiol reactivity. Over seven hundred peptides were detected with modifications but only 9 of 28 annotated active site cysteines in this dataset were modified by F13-IAM. Further exploration of the impacts of 8:2 FTAC adducts on protein function revealed that 8:2 FTAC modification promotes protein aggregation in vitro. These results suggest that PFAS may exhibit significant off-target reactivate and suggests a more general mechanisms of toxicity of PFAS induced protein aggregation.

INSTRUMENT(S): Q Exactive HF-X

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Hui Peng

PROVIDER: MSV000091587 | MassIVE | Wed Mar 29 08:40:00 BST 2023

REPOSITORIES: MassIVE

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