Figure 5.

Reduction of Trx h2r, Trx s1r, and Trx s2r by NTR plus NADPH or other reducing agents. Trxs were incubated in the absence (None) or the presence of DTT (2.5 mm for Trx h2r and Trx s1r or 20 mm for Trx s2r), 5 mm GSH, 0.2 mm NADPH plus 1 μm MtNTRA, or 0.2 mm NADPH alone, during 15 min at RT, after which any SH groups formed during this time were labeled with 5 mm mBBr. Proteins were finally resolved by 15% (w/v) SDS-PAGE and observed under UV light (black segments) before staining with Coomassie Blue (gray segments).