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Description

LC-MS/MS experiment was performed on a 1290 Infinity U-HPLC system (Agilent Technologies, Santa Clara, CA, USA) mounted with a Kinetex Biphenyl column (2.1 mm, 50 mm, 1.7 m, Phenomenex), maintained at 50 C

Mutation of U597 to serine resulted in sustained thiolate-FAD charge transfer absorption and loss of the ability to reduce Trx, indicating that the C596-U597 disulfide functions in the catalytic sequence to receive electrons from the C154 C159 pair and distribute them to Trx