Computational identification of potential dinitroaniline binding sites in kinetoplastid alpha-tubulin

The New Medicines for Trypanosomatidic Infections - NMTrypI project aimed at obtaining new candidate drugs against Trypanosomatidic infections with appropriate efficiency from the lead phase to the final preclinical phase that are more accessible to patients.

Design, synthesis, computational studies and biological evaluation of antiparasitic dinitroaniline-ether phospholipid hybrids

Alignments of various alpha-tubulin and beta-tubulin sequences from dinitroaniline-sensitive and dinitroaniline-resistant species. Sequences were retrieved from UniProt with the identifiers listed below and subjected to a multiple sequence alignment using ClustalOmega (ebi.ac.uk/Tools/msa/clustalo/; ClustalOmega webserver, last accessed 16-02-23):

alpha-tubulin:

• [dinitroaniline sensitive] T. cruzi - Q27352; T. brucei brucei - Q4GYY5; L. infantum - ...

Creation of homology models of various tubulins from dinitroaniline-sensitive and -resistant species, and a comparative analysis of their electrostatic potential grids overall and in putative binding site regions using PIPSA (Protein Interaction Property Similarity Analysis).

Collected PDB-files of homology models of alpha-tubulin, all based on T. thermophila tubulin PDB-ID 5ubq, chain A:

Includes models of the following dinitroaniline herbicide sensitive species (UniProt ID in brackets):

• C. reinhardtii (P09204)
• E. indica (O22347)
• L. donovani (I3W8N6)
• L. infantum (E9AGJ8)
• L. major (Q4QGC5)
• P. falciparum (P14642)
• S. viridis (Q65C79)
• T. brucei (Q4GYY5)
• T. cruzi (Q27352)
• T. gondii (P10873)
• Z. mays (P14640)

Includes models of the following dinitroaniline ...

This archive contains all results of the PIPSA analysis run in zipped form. It contains the following subfolders:

• apbs: Computed electrostatic potential grids, all log files and similarity matrices for the full protein and all sites.
• pdb2pqr: PQR files: conversion results after processing uploaded PDB files with pdb2pqr
• pdbs: Original uploaded PDB files

The following sites have been analyzed (provided are site center (based on T. cruzi model, Ca of the residue) and the sphere radius):

Blume ...

Clustal Omega multiple sequence alignment result for selected alpha-tubulins in FASTA format with gaps. Species and corresponding Uniprot identifiers can be found in the FASTA file.

The following species (Uniprot ID, dinitroaniline-sensitive/resistant) were considered:

• Trypanosoma cruzi (Q27352, sensitive)
• Trypanosoma brucei brucei (Q4GYY5, sensitive)
• Leishmania infantum (E9AGJ8, sensitive)
• Leishmania donovani (I3W8N6, sensitive)
• Leishmania major (Q4QGC5, sensitive)
• Toxoplasma gondii ...

Clustal Omega multiple sequence alignment result for selected beta-tubulins in FASTA format with gaps. Species and corresponding Uniprot identifiers can be found in the FASTA file.

The following species (Uniprot ID, dinitroaniline-sensitive/resistant) were considered:

• Trypanosoma cruzi (Q8STF3, sensitive)
• Trypanosoma brucei brucei (Q4GYY6, sensitive)
• Leishmania infantum (A4HZJ3, sensitive)
• Leishmania donovani (I3W8N7, sensitive)
• Leishmania major (Q4QBZ6, sensitive)
• Toxoplasma gondii ...

General SOP for the webPIPSA workflow to study electrostatic potential differences of full alpha-tubulin homology models and various putative binding site regions

General SOP for alpha-tubulin homology model building

SOP for protein preparation steps performed for alpha-tubulin homology models (hydrogen addition, hydrogen bonding network optimization and energy minimization)