Hrv 3c protease

Pichia pastoris strain KM71H (aox::ARG4, arg4), expression vector pPICZ-A and Zero Blunt TOPO PCR cloning kit were purchased from Invitrogen. A plasmid containing the gene for full-length human STRA6 transcript variant 2 (NM_022369.3) in the pCMV6-XL4 cloning vector, was purchased from OriGene. The pEGFP-N1 plasmid containing eGFP was purchased from Clontech. Zeocin was from Invivogen. DyLight 594 amine-reactive dye was purchased from Pierce. N-dodecyl β-D maltoside (DDM), N-decyl β-D maltoside (DM) and Fos-Choline-12 (FC-12) were from Anatrace. Cholesteryl hemisuccinate, N, N-dimethyldodecylamine N-oxide (LDAO) and dodecyl nonaoxyethylene ether (C12E9) were from Sigma. HRV 3C protease was from Sino Biological. Polyclonal antibody raised against full-length human STRA6 (#H00064220-D01P) was from Abnova. The Biacore 3000 and instrument-specific consumables (CM5 sensor chip, ethyl(dimethylaminopropyl)-carbodiimide, N-hydroxysulphosuccinimide and ethanolamine) were obtained from GE Healthcare.

The GST-IDO1 gene was cloned by Shanghai Generay Biotech Co. Ltd (Shanghai, China). The BL21(DE3) cells were obtained from TransGen Biotech Co. Ltd (Shanghai, China). Ampicillin, isopropyl β-D-thiogalactoside (IPTG), 5-Aminolevulinic acid (5-ALA), phenylmethanesulfonyl fluoride (PMSF), DNase I (from bovine pancreas), reduced glutathione, L-tryptophan, ascorbic acid, methylene blue, catalase, and sodium dithionite were obtained from Sangon Biotech (Shanghai, China) Co. Ltd. DMSO was purchased from Sigma-Aldrich. The CCK8 kit was purchased from Beyotime Biotechnology Co. Ltd.(Shanghai, China). Glutathione agarose beads were purchased from Changzhou Smart-Lifesciences Biotechnology Co. Ltd. (Changzhou, China) PD-10 desalting column was purchased from GE Healthcare Biosciences. HRV 3C Protease was purchased from Sino Biological Inc. (Shanghai, China). Cancer cell lines were purchased from the Cell Bank of Chinese Academy of Sciences. The experimental compounds were provided by Professor Zhao (Shanghai Institute of Organic Sciences, Chinese Academy of Sciences). All chemicals of analytical and reagent grade were obtained from commercial sources and used without further purification.