Poster presentation
37
PROTEIN SEQUENCES ANALYSIS OF THE MAIN MDR PUMP
ACRAB-TOLC FROM ESCHERICHIA COLI:
ANTIBACTERIAL
ACTION OF SUBSTANCES OF PLANT ORIGIN
Marina V. Karakozova,
Vasilisa N. Raldugina
,
Pavel A. Nazarov,
Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State
University, Moscow, Russia
Multidrug-resistance (MDR) pumps are the cornerstone of protecting bacteria from
antibiotics. MDR pumps form the basis of non-specific protection of bacteria and
belong to six families of MDR pumps. One of the best studied MDR pumps is the E.
coli AcrAB-TolC pump. The study of pump sequences
is an important task in
understanding the processes of formation of new clinical isolates-superbugs, since
resistance due to efflux occupies an important place in the total number of resistant
bacteria arising as a result of antibiotic therapy. It is believed
that the genes encoding
MDR pumps are variable and belong to the so-called “luxury” genes, i.e., they are
designed to adapt bacteria to changing environmental conditions. In this work, we
analyzed all known sequences of TolC, AcrA,
and AcrB proteins from
E. coli
strains,
and based on the results obtained, we found important patterns for understanding the
formation of resistance in the bacterial population. TolC, AcrA, and AcrB protein
sequences for
E. coli
strains were downloaded from the NCBI database in genbank and
fasta formats using the biopython package and the Etrez.Esearch and Entrez.Efetch
functions. In this work, all currently existing sequences of the AcrAB-TolC proteins in
the NCBI database were analyzed: TolC (8383 sequences), AcrA (5715 sequences), and
AcrB (3561 sequences). The sequences were aligned with each other using BLASTp
with the creation of an individual database. For further work with the data, the numpy
and pandas packages were used. Analysis of laboratory
strains showed that the
sequences of a large number of laboratory strains used (K-12 and B) are conserved and
do not detect any mutation in the amino acid sequences. This allows us to state that for
each protein there are certain consensus sequences that do not change. Analysis of all
sequences showed that there are several consensus sequences in the
E. coli
population
and for each protein and there are many point mutations around these consensus
sequences. However, consensus sequences
predominate in the population, and this
explains the fact that several decades of laboratory cultivation did not affect laboratory
strains of E. coli. This normalizes the use of different laboratory strains in relation to at
least the AcrAB-TolC pump in various studies. The conservatism of amino acid
sequences of the pumps resembles the conservatism of the “housekeeping” genes,
which indicates the great importance of these genes for bacteria,
despite the fact that
these genes are formally classified as nonessential genes. In this work, we used an
approach to analyze the effect of plant substances based on quinones and terpenes on
their ability to penetrate into bacterial cells.
This research was funded by Russian Science Foundation (RSF) grant 22-15-00099.