title: Expression and functional analysis of novel molecule - Latcripin-13 domain from Lentinula edodes C91-3 produced in prokaryotic expression system.
abstract: The shiitake mushroom Lentinula edodes has health benefits and is used to treat various diseases due to its immunomodulatory and antineoplastic properties. In the present study, the Latcripin-13 domain, isolated from L. edodes, was expressed in Escherichia coli Rosetta-gami(DE3) in the form of inclusion bodies. The Latcripin-13 domain was purified by Ni-His affinity chromatography with high purity and refolded by urea gradient dialysis. The product showed biological activity in A549 cells, a human lung cancer cell line, by flow cytometry and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) method. The MTT assay and the flow cytometry results revealed that there was a great difference between the Latcripin-13 domain-treated group and the control group (p<0.05). Similarly, cell apoptosis observed by transmission electron microscopy (TEM) supported the flow cytometry results. This work demonstrated that the Latcripin-13 domain can induce apoptosis of A549 cells, which will bring new insights into the development of new antitumor drugs in the future.
authors.affiliation: Department of Microbiology, Dalian Medical University, Dalian 116044, China.
authors.forename: Jia
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Microbiology, Dalian Medical University, Dalian 116044, China.
authors.forename: Mintao
authors.initials: M
authors.lastname: Zhong
authors.affiliation: Department of Microbiology, Dalian Medical University, Dalian 116044, China.
authors.forename: Ben
authors.initials: B
authors.lastname: Liu
authors.affiliation: Department of Microbiology, Dalian Medical University, Dalian 116044, China.
authors.forename: Li
authors.initials: L
authors.lastname: Sha
authors.affiliation: Department of Microbiology, Dalian Medical University, Dalian 116044, China.
authors.forename: Yongzhi
authors.initials: Y
authors.lastname: Lun
authors.affiliation: Department of Microbiology, Dalian Medical University, Dalian 116044, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Department of Microbiology, Dalian Medical University, Dalian 116044, China.
authors.forename: Xingyun
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Microbiology, Dalian Medical University, Dalian 116044, China.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Microbiology, Dalian Medical University, Dalian 116044, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Cao
authors.affiliation: Department of Microbiology, Dalian Medical University, Dalian 116044, China.
authors.forename: Anhong
authors.initials: A
authors.lastname: Ning
authors.affiliation: Department of Microbiology, Dalian Medical University, Dalian 116044, China. Electronic address: huangminchao@163.com.
authors.forename: Min
authors.initials: M
authors.lastname: Huang
mesh_terms: D000595:Amino Acid Sequence; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D017209:Apoptosis; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D005434:Flow Cytometry; D005656:Fungal Proteins / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D046529:Microscopy, Electron, Transmission; D008969:Molecular Sequence Data; D009369:Neoplasms / Q000188:drug therapy; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D020942:Shiitake Mushrooms / Q000737:chemistry*
chemical_list: D000970:Antineoplastic Agents; D005656:Fungal Proteins; C000595217:Latcripin-13 protein, Lentinula edodes
keywords: Apoptosis; Expression; Latcripin-13 domain; Lentinula edodes; PHD domain