Identification of a Novel Anti-cancer Protein, FIP-bbo, from Botryobasidium botryosum and Protein Structure Analysis using Molecular Dynamic Simulation

Da Peng Bao,Ying Ying Wu,Hong Yu Chen,Ying Wang,Jun Jun Shang,Ying Nv Gao,Rui Bai

doi:10.1038/s41598-019-42104-1

Da Peng Bao, Ying Ying Wu + Show 5 more

Open Access

https://doi.org/10.1038/s41598-019-42104-1

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Abstract

Fungal immunoregulatory proteins (FIP) are effective small molecule proteins with broad-spectrum immunomodulatory and anti-cancer activities and can be potential agents for the development of clinical drugs and health food additives. In this study, a new member of FIP named FIP-bbo was obtained through Botryobasidium botryosum genome mining. FIP-bbo has the typical characteristics of FIP but is genetically distant from other FIPs. Recombinant FIP-bbo (rFIP-bbo) was produced in an optimized E. coli expression system, and the pure protein was isolated using a Ni-NTA column. Antineoplastic experiments suggested that FIP-bbo is similar to LZ-8 in inhibiting various cancer cells (Hela, Spac-1, and A549) at lower concentrations, but it is not as potent as LZ-8. The molecular mechanism by which FIP-bbo, FIP-fve, and LZ-8 are cytotoxic to cancer cells has been discussed based on molecular dynamics simulation. Point mutations that may improve the thermal stability of FIP-fve and FIP-bbo were predicted. These results not only present a new candidate protein for the development of anticancer adjuvants, but also provide an approach for designing FIPs with high anticancer activity.

Highlights

Fungal immunoregulatory proteins (FIP) have various immunomodulatory activities and are important to researchers
The dendrogram based on the FIP amino acids analysis showed that FIP is mainly divided into three branches (Fig. 1)
The results proved that rFIP-bbo suppressed Hela, Spca-1 and A549 cell migration to the denuded zone, and the migration inhibition effect of rFIP-bbo was slightly lower than that of recombinant LZ-8 (rLZ-8), but higher than that of rFIP-fve

Summary

Introduction

Several experiments showed that FIP is a potent T cell activator that enhances the immune response by modulating the production of important cytokines and molecular factors (IL-2, IFN-γ, IL-1β, TNF-α etc.) in several vertebrate model systems[4]. The ability of these cytokines to produce a tumour suppressing environment indirectly proves that FIP regulates the immune system and has a cytotoxic effect on tumour cells[5]. Rac[1] activity[8] Both rLZ-8 and FIP-fve prevented lung cancer cell proliferation via the increase G1 arrest, their effect strengths are different. The molecular mechanism by which FIP-bbo, FIP-fve, and LZ-8 induce cytotoxicity in cancer cells has been discussed using molecular dynamics simulation

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