Figure 1: Assay of binding activity of AtPDIL1-2 with 2,4,6-TCP. | Scientific Reports

Figure 1: Assay of binding activity of AtPDIL1-2 with 2,4,6-TCP.

From: Disulfide isomerase-like protein AtPDIL1–2 is a good candidate for trichlorophenol phytodetoxification

Figure 1

(A) Expression and purification of the recombinant AtPDIL1-2 in E. coli. The fractions in purification were applied to a 10% SDS-PAGE gel. lane 1, fraction eluted by 250 mm imidazole and 0.1% sucrose monolaurate; lane 2, washing fraction; lane 3, total induced protein; lane 4, molecular mass standards. (B) AtPDIL1-2 catalyzed oxidative folding of reduced denatured RNase on the basis of cCMP hydrolysis, creating a change in the absorbance at 296 nm. (C) E. coli cells that overexpressed the product of AtPDIL1-2 exhibited significant resistance to 2,4,6-TCP. The cells were diluted to 103 cells/μL, and 0.5–10 μL cells were added to the LB plate supplemented with 250 and 500 μM 2,4,6-TCP. (D) Examination of the binding activity of AtPDIL1-2 with 2,4,6-TCP, recombinant AtPDIL1-2 (3.5 μM) was incubated with 50 μM 2,4,6-TCP in 0.5 mL binding buffer [50 mM Tris-HCl buffer (pH 7.0) containing 150 mM NaCl] for 0–3 h at 4 °C. The residual TCP was detected with HPLC with UV detection at 210 nm per 30 min. The x axe was time (minute) and y was mAU (milliabsorbance unit). (E) Effects of TCP on PDI-mediated isomerase activity.

Back to article page