Functional S-persulfidation exploration reveals a novel regulatory mechanism of HNF1α.

Protein S-persulfidation, a post-translational modification mediated by hydrogen sulfide (H2S), plays an essential role in regulating protein function. However, current profiling methods, mainly based on liquid chromatography-tandem mass spectrometry (LC-MS/MS), tend to detect abundant proteins and often miss low-abundance but important regulators like transcription factors (TFs). To better understand the S-persulfidation landscape, we integrated datasets from three species-Homo sapiens, Mus musculus, and Arabidopsis thaliana-and found that TFs are strikingly underrepresented. To address this gap, we applied oxidative proteomics data from the Oximouse database, which reflects age-related redox changes, and identified hepatocyte nuclear factor 1α (HNF1α) as a candidate S-persulfidated TF. We confirmed that H2S induces S-persulfidation at Cys241 of HNF1α, disrupting its DNA binding through structural remodeling. This modification reduced PCSK9 expression, improved lipid profiles, and alleviated atherosclerosis in mice. Our study presents a cross-species strategy to uncover functional S-persulfidation targets and reveals a redox-based transcriptional mechanism involved in lipid metabolism and cardiovascular protection.