title: Identification, characterization and bioactivity of tumor necrosis factor (TNF)-related apoptosis-inducing ligand from Equus caballus.
abstract: Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) is a member of the TNF superfamily and plays multifunctional roles in the immune system. In the present study, a homolog of TRAIL from the Mongolian horse (named ecTRAIL) was identified and characterized. The 870-bp open reading frame encodes a polypeptide of 289 amino acid residues with a predicted molecular weight of 33.47 kDa and pI of 8.47. The genomic structure of ecTRAIL shares a five-exon/six-intron arrangement similar to its orthologs. Multiple alignments show that ecTRAIL is a type II transmembrane protein with a typical transmembrane region, three conserved cysteine residues (Cys56, Cys77, Cys238) and a TNF family signature sequence ([LV]-x-[LIVM]-x(3)-G-[LIVMF]-Y-[LIVMFY](2)-x(2)-[QEKHL]-[LIVMGT]-x-[LIVMFY]). Three-dimensional structure prediction based on the same template revealed that the positional arrangement of the key amino acid residues, Cys238 and Cys230 in ecTRAIL and human TRAIL, respectively, is significantly conserved. Evolutionary analysis suggests that ecTRAIL is most closely related to its ortholog from pigs, with an identity of 83.99%. The solubilizing small ubiquitin-related modifier (SUMO) tag fused recombinant protein SUMO-ecsTRAIL was successfully expressed in E. coli and exhibited binding activity and cytotoxicity to HeLa cells in a cross-species manner in vitro. These results provide a better understanding of TRAILs in mammals and indicate that ecTRAIL may play an important role in the immune response in horses.
authors: Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Sciences College, Nanjing Normal University, Nanjing 210046, People's Republic of China; Qilu Institute of Pharmaceutical Research, Qilu Pharmaceutical Co. Ltd, Jinan 250100, People's Republic of China.
authors: Lei
authors: L
authors: Ma
authors: Co-Innovation Center for Sustainable Forestry in Southern China, Key Laboratory of Forest Genetics & Biotechnology, Ministry of Education, Nanjing Forestry University, Nanjing 210037, People's Republic of China.
authors: Ming
authors: M
authors: Sang
authors: Co-Innovation Center for Sustainable Forestry in Southern China, Key Laboratory of Forest Genetics & Biotechnology, Ministry of Education, Nanjing Forestry University, Nanjing 210037, People's Republic of China. Electronic address: zhangjiaxin60633@163.com.
authors: Jiaxin
authors: J
authors: Zhang
authors: Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Sciences College, Nanjing Normal University, Nanjing 210046, People's Republic of China. Electronic address: zhangshuangquan@njnu.edu.cn.
authors: Shuangquan
authors: S
authors: Zhang
mesh_terms: D000818:Animals; D015478:Antigens, Heterophile / Q000276:immunology; D003001:Cloning, Molecular; D003602:Cytotoxicity, Immunologic; D006367:HeLa Cells; D006736:Horses / Q000276:immunology*; D006801:Humans; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000235:genetics; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D025841:Small Ubiquitin-Related Modifier Proteins / Q000235:genetics; D013552:Swine / Q000276:immunology; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000235:genetics / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D015478:Antigens, Heterophile; D011994:Recombinant Proteins; D025841:Small Ubiquitin-Related Modifier Proteins; D053221:TNF-Related Apoptosis-Inducing Ligand; D014409:Tumor Necrosis Factor-alpha
keywords: Binding activity; Cytotoxicity; Horse; Immune response; Tumor necrosis factor-related apoptosis-inducing ligand