Derivatization-enhanced thiol redox atlas reveals ENO1–SLC7A11 axis as Ferroptosis protector in myocardial injury

Myocardial injury creates intense redox stress with thiol metabolites as molecular redox buffers. Given the limited analytical sensitivity for thiol detection, global detection and functional characterization of thiol networks remain a major bottleneck. Here we develop a derivatization-enhanced mass spectrometry imaging method, assisted by a Python-based annotation platform, to construct a cross-species thiol redox atlas with spatial resolution. We identify 405 thiols, including 170 newly detected in vivo , and show that the endogenous thiol pool is highly tissue- and region-specific. In the isoproterenol-induced rat model of myocardial injury, heart damage causes distinctive thiol metabolism patterns, especially previously overlooked noncanonical cysteine-derived, long-chain, and substituted thiols. Similar thiol shifts are observed in serum from patients with myocardial infarction. Upon myocardial injury, multi-omics integration highlights enolase 1 (ENO1) as a thiol-linked hub in cardiomyocytes. ENO1 inhibition weakens SLC7A11 induction, depletes glutathione, and aggravates iron-dependent lipid damage. Biochemical assays support a stress-inducible ENO1–SLC7A11 interaction that promotes cysteine-driven thiol replenishment, highlighting a potential mechanism for cardioprotection. A MS-based strategy mapped 405 endogenous thiols, including 170 newly identified species, and uncovered an ENO1–SLC7A11 axis that suppresses ferroptosis, highlighting promising therapeutic targets for cardioprotection.