Screening of nitroanilines for their affinity to miscanthus α-tubulin (Q70ZL7) for further in vitro polyploidization of miscanthus species

  • О. В. Мельничук
  • С. П. Ожерєдов
  • Д. Б. Рахметов
  • А. І. Ємець
  • Я. Б. Блюм

Abstract

Aim. Biofuel is one of the most prominent directions in contemporary energy technologies. In many cases polyploids are superior to their original forms. Creation of polyploids of miscanthus species may allow obtaining of great amount of high quality biomass. Wide range of antimitotic agents can be used to block formation of spindle fiber during cell division. However, most of them are highly toxic for plant cells. Dinitroanilines are known to be highly efficient in preventing of microtubules to be assembled. The aim of the study was to screen wide range of nitroanilines for their affinity to miscanthus α-tubulin and, thus, to identify the most promising to be applied in polyploidization of miscanthus species. Methods. Derivatives of nitro- and dinitroanilines have been used in the study. On line tools have been applied to find sequences of miscanthus α-tubulines with the aid of database of protein sequences UniProt. Reconstruction of spatial structure of miscanthus α-tubulin has been done in silico by modelling based on homology. Matrix structures have been obtained from data base RCSB Protein Data Bank. The results have been verified with the aid of service MolProbity. Binding of dinitroanilines to α‑tubulin molecules has been analyzed with the aid of CCDC GOLD software. Results. Protein sequence of Miscanthus sinensis α-tubulin (Q70ZL7) has been chosen from 14 found and used in the study. Spatial structure of the molecule has been reconstructed and verified. Two nitroanilines docking sites have been identified and analyzed on the surface of the α-tubulin molecule. Several compounds have been determined to be the most efficient to form ligand-protein complex with α‑tubulin (Q70ZL7) based on GoldScore and ChemScore estimation. Conclusions. Reconstruction and verification of spatial structure of miscanthus α-tubulin molecule have been conducted. Ability of both new and classic dinitroanilines to form ligand-protein complex with α‑tubulin of miscanthus has been evaluated. Four compounds were found to be the most promising for further research on in vitro polyploidization of miscanthus species.

Keywords: nitroanilines, α-tubulin, docking sites, polyploidization, Miscanthus spp.