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. 2009 Sep 23;284(47):32425–32433. doi: 10.1074/jbc.M109.047092

FIGURE 3.

FIGURE 3.

rhMIF was reacted with a 10-fold excess of PEITC for 10 min before analysis of PEITC-adduct formation by mass spectrometry. A, shown is wild type rhMIF before (i) and after (ii) incubation with PEITC. A peak with a mass of 12,508 Da, corresponding to MIF (12,345 Da) with the addition of 1 PEITC (163 Da) resulted. In addition, a peak with a mass of 12,248 Da was also apparent. B, shown is rhMIF with a C60S mutation (i), a C57S mutation (ii) or a C81S mutation (iii) (12329 Da) after incubation with PEITC. In all cases, a peak with a mass of 12,492 Da, corresponding to cysteine mutant MIF with addition of one PEITC, resulted. In addition, a peak with a mass of 12,232 Da was also formed. C, shown is P2A rhMIF before (i) and after (ii) incubation with PEITC. Only the mass of the parent protein (12,319 Da) was observed.