Identification and Functional Characterization of a Novel Immunomodulatory Protein From Morchella conica SH.

Abstract

A novel fungal immunomodulatory protein (FIP) was found in the precious medical and edible mushroom Morchella conica SH, defined as FIP-mco, which belongs to the FIP family. Phylogenetic analyses of FIPs from different origins were performed using Neighbor-Joining method. It was found that FIP-mco belonged to a new branch of the FIP family and may evolved from a different ancestor compared with most other FIPs. The cDNA sequence of FIP-mco was cloned and expressed in the yeast Pichia Pastoris X33. The recombinant protein of FIP-mco (rFIP-mco) was purified by agarose Ni chromatography and determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis. The protein rFIP-mco could significantly suppress the proliferation of A549 and HepG2 cells at the concentration of 15 and 5 μg/ml, respectively, and inhibited the migration and invasion of human A549 and HepG2 cells at the concentration of 15 and 30 μg/ml respectively in vitro. Further, rFIP-mco can significantly reduce the expression levels of TNF-α, IL-1β, and IL-6 in the THP1 cells (human myeloid leukemia mononuclear cells). In order to explore the potential mechanism of the cytotoxicity effect of rFIP-mco on A549 and HepG2 cells, cell cycle and apoptosis assay in the two cancer cells were conducted. The results demonstrated that G0/G1 to S-phase arrest and increased apoptosis may contribute to the proliferation inhibition by rFIP-mco in the two cancer cells. Molecular mechanism of rFIP-mco's reduction effect on the inflammatory cytokines was also studied by suppression of the NF-κB signaling pathway. It showed that suppression of NF-κB signaling is responsible for the reduction of inflammatory cytokines by rFIP-mco. The results indicated the prospect of FIP-mco from M. conica SH as an effective and feasible source for cancer therapeutic studies and medical applications.

• Authors: Guogan Wu, Yu Sun, Tingshan Deng, Lili Song, Peng Li, Haijuan Zeng, Xueming Tang
• Journal: Frontiers in immunology (Front Immunol) Vol. 11 Pg. 559770 ( 2020) ISSN: 1664-3224 [Electronic] Switzerland
• PMID: 33193329 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2020 Wu, Sun, Deng, Song, Li, Zeng and Tang.

Chemical References

• Cytokines
• Fungal Proteins
• Inflammation Mediators
• NF-kappa B
• Recombinant Proteins

Topics

• Amino Acid Sequence
• Apoptosis (drug effects)
• Ascomycota (classification, genetics, immunology, metabolism)
• Cell Cycle (drug effects)
• Cell Line
• Cell Movement (drug effects, immunology)
• Cell Proliferation (drug effects)
• Computational Biology (methods)
• Cytokines (metabolism)
• Databases, Genetic
• Fungal Proteins (chemistry, genetics, metabolism, pharmacology)
• Humans
• Immunomodulation (drug effects)
• Inflammation Mediators (metabolism)
• NF-kappa B (metabolism)
• Phylogeny
• Recombinant Proteins (genetics, metabolism, pharmacology)
• Signal Transduction