Use of a molybdenum(VI) complex as artificial protease in protein photocleavage

• This is the first demonstration of protein photocleavage by a Mo(VI) complex.
• The specific cleavage reaction undergoes at room temperature, pH 7 under UV light.
• The cleavage starts to occur at 10 min of irradiation.
• Hydroxyl radicals may be involved in the cleavage reaction.

In this study, a molybdenum(VI) peroxo α-amino acid complex, MoO(O2)2(α-leucine) (H2O), was prepared and used as an artificial protease for site-specific cleavage of porcine pepsin, a model protein. Cleavage of pepsin by MoO(O2)2(α-leucine) (H2O) was achieved under photochemical conditions at room temperature and pH 7.0. The reaction was activated by irradiation of the MoO(O2)2(α-leucine) (H2O)-protein mixture by UV light (320 and 340 nm) for up to 30 min. No cleavage was observed in the absence of MoO(O2)2(α-leucine) (H2O) or the light. The photocleavage yield increased with irradiation time. The cleaved fragments were sequencable, and the cleavage site was assigned to Leu(112)–Tyr(113). The cleavage reaction was quenched by ethanol. Therefore, hydroxyl radicals may be involved in the reaction and responsible for the cleavage of the protein. This is the first demonstration of the successful photocleavage of proteins by a molybdenum complex. This observation can provide a new approach for the photochemical footprinting of metal binding sites on proteins.

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