Querying the In Vitro Proteome Cysteine Reactivity of 8:2 Fluorotelomer Acrylate.
David Ross HallJeremy R GauthierHui PengPublished in: Environmental science & technology (2023)
Despite the phase out of legacy per- and polyfluoroalkyl substances (PFAS), fluorotelomer-based polymers (FTP) have been used for many applications, notably textile surface coatings. FTPs are of a health concern 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 evaluate this hypothesis, 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 bottom-up data-dependent proteomics analysis. Using this method, over 100 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 C 6 F 13 -iodoacetamide (F 13 -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 F 13 -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 8:2 FTAC may exhibit significant proteome thiol reactivity and imply a more general mechanism of toxicity of PFAS-induced protein aggregation.
Keyphrases
- amino acid
- liquid chromatography
- solid phase extraction
- protein protein
- mass spectrometry
- binding protein
- public health
- living cells
- high performance liquid chromatography
- simultaneous determination
- small molecule
- mental health
- electronic health record
- fluorescent probe
- molecularly imprinted
- climate change
- drug induced
- high resolution
- high glucose
- gas chromatography