Figure 6

Analysis of mature human resistin expressed in the cytoplasm of E. coli at 30°C in LB. Panel A) Coomassie stained SDS-PAGE analysis of purified MBPx-resistin indicates production of the disulfide linked dimer increases with pre-expression of Erv1p and DsbC in both wild-type and Δgor ΔtrxB strains. The molecular weights (in kDa) of the markers (mw) are indicated as are the positions of the monomeric and dimeric forms of MBPx-resistin. Panel B) Coomassie stained SDS-PAGE analysis of mature resistin produced in origami with pre-expression of Erv1p and DsbC. S = soluble fraction of E. coli lysates; P = Purified MBPx-resistin from an amylose column; D = Purified material that has been digested with factor Xa to release MBP and resistin (marked with *). Note that under non-reducing conditions the resistin is a disulfide linked homo-dimer. The molecular weights (in kDa) of the markers (mw) are indicated as is the position of MBP post digestion. The position of the MBP-resistin fusion is indicated with an arrow.