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. 2012 Dec 7;288(5):3346–3358. doi: 10.1074/jbc.M112.418046

FIGURE 6.

FIGURE 6.

hTrx1-Cys-73 bridges rtTF-Cys-209. The mixture of hTrx1 and rtTF was incubated with IAM to block free thiols and then separated by non-reducing SDS-PAGE. The protein band corresponding to TF-hTrx1 complex was cut out for in-gel tryptic digestion and LC-ESI-MS/MS analysis. A, MS/MS signals showed those corresponding to IAM-labeled Cys-47, Cys-59, and Cys-189 from rtTF. MS signals revealed a fragment containing Cys-47/-59 disulfide of rtTF. A peptide containing rtTF-Cys-209 was missing. B, MS/MS signals showed those corresponding to IAM-labeled Cys-62 and Cys-69 from hTrx1. MS signals contained a fragment containing Cys-32/-35 disulfide of hTrx1. A peptide containing Cys-73 was missing.