title: Preparation and characterization of a novel variant of human tumor necrosis factor-related apoptosis-inducing ligand from the rhesus monkey, Macaca mulatta.
abstract: Human tumor necrosis factor-related apoptosis-inducing ligand (hTRAIL) and its variants are attractive antitumor drug candidates. The predicted amino acid sequence of the functional extracellular domain of Macaca mulatta TRAIL (mmTRAIL) was found to differ from that of hTRAIL at four positions. In this study, the gene encoding mmTRAIL was cloned and recombinantly expressed in Escherichia coli at a yield of approximately 20-30 mg/L, which was two times higher than that of hTRAIL. SDS-PAGE showed that denatured mmTRAIL and hTRAIL had similar molecular weights. However, size-exclusion chromatography and dynamic light scattering (DLS) analysis demonstrated that the molecular size of native mmTRAIL was smaller than that of native hTRAIL. Cooling solutions of these proteins from room temperature to 0 °C induced considerable precipitation of hTRAIL but not of mmTRAIL, indicating that mmTRAIL was more soluble than hTRAIL at low temperatures. Additionally, mmTRAIL was more resistant than hTRAIL to N-bromosuccinimide (NBS)-induced precipitation. Although mmTRAIL and hTRAIL showed comparable nanomolar affinities for human death receptors, the dissociation rate of the mmTRAIL-receptor complex was slower than that of the hTRAIL-receptor complex, suggesting that the mmTRAIL-receptor complex was more stable. Moreover, mmTRAIL induced caspase-dependent apoptosis in human tumor cells with an IC50 that was two to three times lower than that of hTRAIL. However, in vivo evaluation demonstrated that mmTRAIL or hTRAIL led to a similar level of tumor suppression in mice bearing COLO205 xenografts. Nevertheless, the advantage of its better solubility should promote the production and further use of mmTRAIL in cancer biotherapy.
authors.affiliation: Key Lab of Transplant Engineering and Immunology, MOH, West China Hospital, Sichuan University, Chengdu, 610041, China.|Regenerative Medical Research Center, West China Hospital, Sichuan University, Chengdu, 610041, China.
authors.forename: Dianlong
authors.initials: D
authors.lastname: Jia
authors.affiliation: Key Lab of Transplant Engineering and Immunology, MOH, West China Hospital, Sichuan University, Chengdu, 610041, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Yang
authors.affiliation: Key Lab of Transplant Engineering and Immunology, MOH, West China Hospital, Sichuan University, Chengdu, 610041, China.
authors.forename: Ze
authors.initials: Z
authors.lastname: Tao
authors.affiliation: Key Lab of Transplant Engineering and Immunology, MOH, West China Hospital, Sichuan University, Chengdu, 610041, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Wan
authors.affiliation: Key Lab of Transplant Engineering and Immunology, MOH, West China Hospital, Sichuan University, Chengdu, 610041, China.
authors.forename: Jingqiu
authors.initials: J
authors.lastname: Cheng
authors.affiliation: Key Lab of Transplant Engineering and Immunology, MOH, West China Hospital, Sichuan University, Chengdu, 610041, China. xiaofenglu@yahoo.com.
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Lu
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D001974:Bromosuccinimide / Q000737:chemistry; D020169:Caspases / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D011232:Chemical Precipitation; D003001:Cloning, Molecular; D003110:Colonic Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008253:Macaca mulatta; D051379:Mice; D008970:Molecular Weight; D000072417:Protein Domains; D053218:Receptors, Death Domain / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D012995:Solubility; D013045:Species Specificity; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D023041:Xenograft Model Antitumor Assays
chemical_list: D000970:Antineoplastic Agents; D053218:Receptors, Death Domain; D011994:Recombinant Proteins; D053221:TNF-Related Apoptosis-Inducing Ligand; C505653:TNFSF10 protein, human; D020169:Caspases; D001974:Bromosuccinimide
keywords: Anticancer drug; Apoptosis; Cancer biotherapy; Rhesus monkey; Tumor necrosis factor-related apoptosis-inducing ligand