Expression and functional analysis of novel molecule - Latcripin-13 domain from Lentinula edodes C91-3 produced in prokaryotic expression system

Gene. 2015 Jan 25;555(2):469-75. doi: 10.1016/j.gene.2014.11.014. Epub 2014 Nov 12.

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.

Keywords: Apoptosis; Expression; Latcripin-13 domain; Lentinula edodes; PHD domain.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Antineoplastic Agents / chemistry
  • Antineoplastic Agents / pharmacology*
  • Apoptosis
  • Cell Line, Tumor
  • Cloning, Molecular
  • Flow Cytometry
  • Fungal Proteins / chemistry
  • Fungal Proteins / pharmacology*
  • Humans
  • Microscopy, Electron, Transmission
  • Molecular Sequence Data
  • Neoplasms / drug therapy
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Shiitake Mushrooms / chemistry*

Substances

  • Antineoplastic Agents
  • Fungal Proteins
  • Latcripin-13 protein, Lentinula edodes