Disulfide-Centered Hydrogen Bonding: Insights from Protein Structure Analysis and IR–UV Double Resonance Spectroscopy

Despite the importance of disulfide bonds in proteins, direct experimental characterization of noncovalent interactions involving their sulfur atoms, particularly sulfur-centered hydrogen bonds (H-bonds), remains underexplored. Here, we present an integrated study combining Protein Data Bank (PDB) analysis, quantum chemical calculations, and gas-phase vibrational spectroscopy. PDB screening revealed disulfide bonds in ∼20% of protein structures, with nearly 20 000 potential O–H···S and N–H···S H-bonds. The O–H···S H-bonds are shorter and more directional than the N–H···S H-bonds, consistent with the topological and energetic analyses of model systems. Mass-selective electronic and IR spectroscopy on the jet-cooled p -cresol–dimethyl disulfide (pCR–DMDS) complex confirmed disulfide-centered H-bonds through red-shifts in S 1 → S 0 electronic transitions and O–H stretching frequencies (Δν O–H ). The experimental Δν O–H shift and comparative analysis with other H-bond acceptors (H 2 S, H 2 O, and dimethylsulfide) provide benchmark data on the intrinsic strength of disulfide-centered H-bonds, crucial for refining computational models and enhancing the understanding of their role in protein structure and function.