A NEW APPROACH IN THE DEVELOPMENT OF A DENTAL CARIES VACCINE S. P. Rubnikovich, V. V. Poboinev, V. V. Khrustalev Proceedings of the National Academy of Sciences of Belarus Medical Series, 2024 The genomes of the bacteria Streptococcus mutans and Streptococcus sobrinus, which cause dental caries, currently have been fully sequenced. However, the secondary and tertiary structures of the full-size surface proteins of these microorganisms, by which they attach to the surface of teeth covered with saliva, have not been instrumentally determined at the moment. There are a number of experimental studies on the use of these proteins in the process of developing a dental caries vaccine. However, there is currently no commercially available dental caries vaccine.The aim of the study was to choose an antigen for subsequent molecular modeling of a unique peptide for the development of a dental caries vaccine.To develop an effective and safe dental caries vaccine, it is necessary to perform a number of experiments in silico, preceding experiments in vitro and in vivo. Today, this approach is not only generally recognized, but also allows to significantly reduce the cost of experiments and time at the preclinical and clinical studies. According to our hypothesis, as an antigen for the development of a dental caries vaccine, it is necessary to use a short fragment of the surface protein (a peptide) of Streptococcus mutans and/or Streptococcus sobrinus, whose homology in amino acid sequence is 84.8 %, the spatial structure of which should correspond to the spatial structure of the corresponding fragment in a full-sized protein. In addition, the selected protein fragment, which will be part of the vaccine peptide, must be available to antibodies, i. e. located on the surface of the protein and defined as a B-cell linear and spatial epitope. Also, according to our hypothesis, the vaccine peptide may consist of the most stable fragments of alanine and proline rich regions of the surface protein of Streptococcus mutans and/or Streptococcus sobrinus for mutual stabilization of the spatial structure.
Structural Shifts of the Parvovirus B19 Capsid Receptor-binding Domain: A Peptide Study Vladislav Victorovich Khrustalev, Aleksander Nicolaevich Stojarov, Anastasia Aleksandrovna Akunevich, Oleg Evgenyevich Baranov, Anna Vladimirovna Popinako, et al. Protein and Peptide Letters, 2024 Background:: Binding appropriate cellular receptors is a crucial step of a lifecycle for any virus. Structure of receptor-binding domain for a viral surface protein has to be determined before the start of future drug design projects. Objective:: Investigation of pH-induced changes in the secondary structure for a capsid peptide with loss of function mutation can shed some light on the mechanism of entrance. Methods:: Spectroscopic methods were accompanied by electrophoresis, ultrafiltration, and computational biochemistry. Results:: In this study, we showed that a peptide from the receptor-binding domain of Parvovirus B19 VP1 capsid (residues 13-31) is beta-structural at pH=7.4 in 0.01 M phosphate buffer, but alpha- helical at pH=5.0, according to the circular dichroism (CD) spectroscopy results. Results of infra- red (IR) spectroscopy showed that the same peptide exists in both alpha-helical and beta-structural conformations in partial dehydration conditions both at pH=7.4 and pH=5.0. In contrast, the peptide with Y20W mutation, which is known to block the internalization of the virus, forms mostly alpha-helical conformation in partial dehydration conditions at pH=7.4. According to our hypothesis, an intermolecular antiparallel beta structure formed by the wild-type peptide in its tetramers at pH=7.4 is the prototype of the similar intermolecular antiparallel beta structure formed by the corresponding part of Parvovirus B19 receptor-binding domain with its cellular receptor (AXL). Conclusion:: Loss of function Y20W substitution in VP1 capsid protein prevents the shift into the beta-structural state by way of alpha helix stabilization and the decrease of its ability to turn into the disordered state.
Equilibrium Between Dimeric and Monomeric Forms of Human Epidermal Growth Factor is Shifted Towards Dimers in a Solution Anastasia Aleksandrovna Akunevich, Vladislav Victorovich Khrustalev, Tatyana Aleksandrovna Khrustaleva, Victor Vitoldovich Poboinev, Nikolai Vladimirovich Shalygo, et al. Protein Journal, 2022 An interplay between monomeric and dimeric forms of human epidermal growth factor (EGF) affecting its interaction with EGF receptor (EGFR) is poorly understood. While EGF dimeric structure was resolved at pH 8.1, the possibility of EGF dimerization under physiological conditions is still unclear. This study aimed to describe the oligomeric state of EGF in a solution at physiological pH value. With centrifugal ultrafiltration followed by blue native gel electrophoresis, we showed that synthetic human EGF in a solution at a concentration of 0.1 mg/ml exists mainly in the dimeric form at pH 7.4 and temperature of 37 °C, although a small fraction of its monomers was also observed. Based on bioinformatics predictions, we introduced the D46G substitution to examine if EGF C-terminal part is directly involved in the intermolecular interface formation of the observed dimers. We found a reduced ability of the resulting EGF D46G dimers to dissociate at temperatures up to 50 °C. The D46G substitution also increased the intermolecular antiparallel β-structure content within the EGF peptide in a solution according to the CD spectra analysis that was confirmed by HATR-FTIR results. Additionally, the energy transfer between Tyr and Trp residues was detected by fluorescence spectroscopy for the EGF D46G mutant, but not for the native EGF. This allowed us to suggest the elongation and rearrangement of the intermolecular β-structure that leads to the observed stabilization of EGF D46G dimers. The results imply EGF dimerization under physiological pH value and temperature and the involvement of EGF C-terminal part in this process.
The cytoplasmic tail of influenza A/H1N1 virus hemagglutinin is β-structural Vladislav V. Khrustalev, Larisa V. Kordyukova, Alexander M. Arutyunyan, Victor V. Poboinev, Tatyana A. Khrustaleva, et al. Journal of Biomolecular Structure and Dynamics, 2022 Influenza A/H1N1 virus hemagglutinin (HA) is an integral type I glycoprotein that contains a large glycosylated ectodomain, a transmembrane domain, and a cytoplasmic tail (CT) of 10-14 amino acid residues. There are absolutely no data on the secondary or tertiary structure of the HA CT, which is important for virus pathogenesis. Three highly conserved cysteines are post-translationally modified by the attachment of fatty acid residues that pin the CT to the lipid membrane inside the virion. We applied circular dichroism (CD) and fluorescence spectroscopy analysis to examine four synthetic peptides corresponding to 14-15 C-terminal residues of H1 subtype HA (NH2-WMCSNGSLQCRICI-COOH; NH2-FWMCSNGSLQCRICI-COOH), with free or acetaminomethylated cysteines, in the reduced or non-reduced state, at various pH values and temperatures. The CD analysis detected the formation of a β-structure (30-65% according to the new BeStSel algorithm), in addition to an unstructured random coil, in every peptide in various conditions. It was completely or partially recognized as an antiparallel β-structure that was also confirmed by the multi-bounce Horizontal Attenuated Total Reflectance Fourier Transformed Infrared (HATR-FTIR) spectroscopy analysis. According to the experimental data, as well as 3 D modeling, we assume that the amino acid sequence corresponding to the HA CT may form a short antiparallel β-structure under the lipid membrane within a virion. Communicated by Ramaswamy H. Sarma.
Spectra of tryptophan fluorescence are the result of co-existence of certain most abundant stabilized excited state and certain most abundant destabilized excited state Khrustalev Vladislav Victorovich, Khrustaleva Tatyana Aleksandrovna, Poboinev Victor Vitoldovich, Stojarov Aleksander Nicolaevich, Kordyukova Larisa Valentinovna, et al. Spectrochimica Acta Part A Molecular and Biomolecular Spectroscopy, 2021 Fluorescence spectra of proteins and peptides are traditionally used to get an information on self-association of proteins and peptides, on their tertiary and quaternary structure. In this study it was shown that there are just three peaks of tryptophan fluorescence (at ∼308, at ∼330, and at ∼360 nm) in rough unsmoothed spectra of fluorescence of pure tryptophan in different solvents that change their heights depending on the polarity of a solvent. Two separate peaks at ∼330 nm and ∼360 nm are especially prominent in the spectrum of human epidermal growth factor. In contrast, in smoothed (either mathematically, or physically) spectra of Trp-containing proteins a single maximum of fluorescence varies between 330 and 360 nm. The theory of tryptophan fluorescence is discussed in light of three discrete peaks existence. A stabilizing hydrogen bond with aromatic system of benzene ring in the excited state is proposed as the cause of emission at ∼360 nm bringing Trp to the destabilized ground state. Emission from the destabilized excited state has a maximum at ∼330 nm if the ground state is destabilized, as well as if both states are stabilized. If the excited state is destabilized, while the ground state is stabilized by purely hydrophobic interactions, emitted light should have a maximum at ∼308 nm. The degree of hydrophilicity of tryptophan microenvironment is proposed to be measured as the ratio between the peak at 360 nm and the peak at 330 nm if the observed shifts are not "horizontal", but "vertical". The process of dissociation of hemagglutinin trimers from pandemic Influenza A(H1N1) virus is described as an example of the advantages of the proposed method.
Influence of mutations caused by radiation exposure on the bilirubin binding sites of human serum albumin V. V. Poboinev, V. V. Khrustalev, A. N. Stojarov, T. A. Khrustaleva Proceedings of the National Academy of Sciences of Belarus Medical Series, 2021 In this article we analyze the bilirubin binding sites of human serum albumin from the point of view of the secondary structure instability, as well as the effect of amino acid substitutions caused by radiation exposure on the ability of albumin to bind bilirubin-IX-alpha. Based on calculations of binding energy and inhibition constants of bilirubin-albumin complexes before and after the amino acid substitutions, it was found that amino acid substitutions have different effects on the ability of human serum albumin to bind bilirubin. Amino acid substitutions Asp269-Gly269 (Nagasaki-1), Glu354-Lys354 (Hiroshima-1), Asp375-Asn375 (Nagasaki-2) reduce the binding free energy of bilirubin with human serum albumin, and the amino acid substitutions His3-Gln3 (Nagasaki-3) and Glu382-Lys382 (Hiroshima-2) increase it during molecular docking with the corresponding areas of the protein surface. The inhibition constants are significantly higher than with known binding sites. In general, mutations caused by radiation exposure cannot effect on bilirubin binding sites of human serum albumin, since the amino acid residues that are replaced do not interact with the amino acid residues from the binding sites (Leu115, Arg117, Phe134, Tyr138, Ile142, Phe149, Phe157, Tyr161, Arg186, Lys190, Lys240, Arg222). All amino acid residues from known binding sites are located in stable elements of the secondary structure of human serum albumin.The data obtained are important for understanding the impact of radiation exposure on the development of bilirubin encephalopathy in the population of the Chernobyl region and Japan.
Взаимодействия эндогенных лигандов с сывороточным альбумином человека ВВ Побойнев, АН Стожаров Доклады Национальной академии наук Беларуси 70 (2), 141-152 , 2026 2026
Conjugation with the Carrier Helped to Reveal acidification-Induced Structural Shift in the Peptide from Phospholipase Domain of Parvovirus B19 VV Khrustalev, OV Khrustaleva, AN Stojarov, AA Akunevich, OE Baranov, ... The Protein Journal 43 (4), 805-818 , 2024 2024
Новый подход к разработке вакцины от кариеса зубов СП Рубникович, ВВ Побойнев, ВВ Хрусталёв Известия Национальной академии наук Беларуси. Серия медицинских наук 21 (2 … , 2024 2024
Random coil to alpha helix structural shift in the peptide from phospholipase domain of Parvovirus B19 capsid conjugated with the carrier is caused by acidification of the medium VV Khrustalev, OV Khrustaleva, AN Stojarov, AA Akunevich, OE Baranov, ... 2024
Structural shifts of the Parvovirus B19 capsid receptor-binding domain: a peptide study VV Khrustalev, AN Stojarov, AA Akunevich, OE Baranov, AV Popinako, ... Protein and Peptide Letters 31 (2), 128-140 , 2024 2024 Citations: 2
Peptide models of the cytoplasmic tail of influenza A/H1N1 virus hemagglutinin expand understanding its pH-dependent modes of interaction with matrix protein M1 VV Poboinev, VV Khrustalev, AA Akunevich, NV Shalygo, AN Stojarov, ... The Protein Journal 42 (4), 288-304 , 2023 2023 Citations: 3
ОЦЕНКА СТУДЕНТАМИ ПЕРВОГО КУРСА ПЕДИАТРИЧЕСКОГО ФАКУЛЬТЕТА УЧЕБНОГО ПРОЦЕССА НА КАФЕДРЕ ОБЩЕЙ ХИМИИ УЧРЕЖДЕНИЯ ОБРАЗОВАНИЯ «БЕЛОРУССКИЙ ГОСУДАРСТВЕННЫЙ МЕДИЦИНСКИЙ УНИВЕРСИТЕТ» ВВ Побойнев, ВВ Хрусталёв, АР Козел, ТВ Латушко, ТВ Прохорова Актуальные проблемы довузовской подготовки, 23-27 , 2023 2023
Лабораторные работы по медицинской химии ВВ Хрусталёв, ТВ Латушко, ТВ Прохорова, ВВ Побойнев, АА Акуневич, ... БГМУ , 2023 2023
The PentUnFOLD algorithm as a tool to distinguish the dark and the light sides of the structural instability of proteins VV Poboinev, VV Khrustalev, TA Khrustaleva, TE Kasko, VD Popkov Amino Acids 54 (8), 1155-1171 , 2022 2022 Citations: 12
The cytoplasmic tail of influenza A/H1N1 virus hemagglutinin is β-structural VV Khrustalev, LV Kordyukova, AM Arutyunyan, VV Poboinev, ... Journal of Biomolecular Structure and Dynamics 40 (10), 4642-4661 , 2022 2022 Citations: 10
Equilibrium between dimeric and monomeric forms of human epidermal growth factor is shifted towards dimers in a solution AA Akunevich, VV Khrustalev, TA Khrustaleva, VV Poboinev, NV Shalygo, ... The Protein Journal 41 (2), 245-259 , 2022 2022 Citations: 3
Научный и образовательный процесс на кафедре общей химии БГМУ в условиях глобальных вызовов и цифровизации ТВ Прохорова Могилевский государственный университет имени АА Кулешова , 2022 2022 Citations: 1
О ВЛИЯНИИ АМИНОКИСЛОТНОЙ ЗАМЕНЫ Y20W НА ВТОРИЧНУЮ СТРУКТУРУ ПЕПТИДА, СООТВЕТСТВУЮЩЕГО ФРАГМЕНТУ РЕЦЕПТОР-СВЯЗЫВАЮЩЕГО ДОМЕНА ПАРВОВИРУСА В19 ВВ Хрусталёв, ТА Хрусталёва, НВ Шалыго, АН Стожаров, ЕГ Сапон, ... Молекулярные, мембранные и клеточные основы функционирования биосистем. К … , 2022 2022
Инновационные технологии при изучении медицинской химии ТВ Прохорова, ТВ Латушко, ВВ Побойнев ББК 28.073 я431 С 23, 209 , 2022 2022 Citations: 3
ОСОБЕННОСТИ КОЙЛА, СПОСОБНОГО К СТРУКТУРНОМУ ПЕРЕХОДУ В ПОЛНОСТЬЮ НЕУПОРЯДОЧЕННОЕ СОСТОЯНИЕ ВВ Побойнев, ВВ Хрусталёв Современные проблемы медицинской биохимии, 219-223 , 2022 2022
СТРУКТУРНЫЕ ОСОБЕННОСТИ САЙТОВ СВЯЗЫВАНИЯ ХЛОРИД-ИОНОВ БЕЛКАМИ БАКТЕРИЙ С РАЗНОЙ GC-НАСЫЩЕННОСТЬЮ ГЕНОМОВ ЮА Бондарь, ВВ Побойнев, ВВ Хрусталёв, ЕГ Ясевич " БИОХИМИЯ XXI ВЕКА" 26 ноября 2021 года, 52 , 2021 2021
Spectra of tryptophan fluorescence are the result of co-existence of certain most abundant stabilized excited state and certain most abundant destabilized excited state KV Victorovich, KT Aleksandrovna, PV Vitoldovich, SA Nicolaevich, ... Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 257, 119784 , 2021 2021 Citations: 45
Analysis of helix-to-coil transitions in human serum albumin V Paboineu, V Khrustalev, A Stazharau, T Khrustaleva FEBS OPEN BIO 11, 172-172 , 2021 2021
MOST CITED SCHOLAR PUBLICATIONS
Spectra of tryptophan fluorescence are the result of co-existence of certain most abundant stabilized excited state and certain most abundant destabilized excited state KV Victorovich, KT Aleksandrovna, PV Vitoldovich, SA Nicolaevich, ... Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 257, 119784 , 2021 2021 Citations: 45
Translation-associated mutational U-pressure in the first ORF of SARS-CoV-2 and other coronaviruses VV Khrustalev, R Giri, TA Khrustaleva, SK Kapuganti, AN Stojarov, ... Frontiers in microbiology 11, 559165 , 2020 2020 Citations: 23
Microenvironment of tryptophan residues in proteins of four structural classes: applications for fluorescence and circular dichroism spectroscopy VV Khrustalev, VV Poboinev, AN Stojarov, TA Khrustaleva European Biophysics Journal 48 (6), 523-537 , 2019 2019 Citations: 22
Amino acid content of beta strands and alpha helices depends on their flanking secondary structure elements VV Khrustalev, TA Khrustaleva, VV Poboinev Biosystems 168, 45-54 , 2018 2018 Citations: 19
Cobalt( ii ) cation binding by proteins VV Khrustalev, TA Khrustaleva, VV Poboinev, CI Karchevskaya, ... Metallomics 11 (10), 1743-1752 , 2019 2019 Citations: 17
The PentUnFOLD algorithm as a tool to distinguish the dark and the light sides of the structural instability of proteins VV Poboinev, VV Khrustalev, TA Khrustaleva, TE Kasko, VD Popkov Amino Acids 54 (8), 1155-1171 , 2022 2022 Citations: 12
The part of a long beta hairpin from the scrapie form of the human prion protein is reconstructed in the synthetic CC36 protein VV Khrustalev, TA Khrustaleva, K Szpotkowski, VV Poboinev, ... Proteins: Structure, Function, and Bioinformatics 84 (10), 1462-1479 , 2016 2016 Citations: 11
The cytoplasmic tail of influenza A/H1N1 virus hemagglutinin is β-structural VV Khrustalev, LV Kordyukova, AM Arutyunyan, VV Poboinev, ... Journal of Biomolecular Structure and Dynamics 40 (10), 4642-4661 , 2022 2022 Citations: 10
Mutational pressure and natural selection in epidermal growth factor receptor gene during germline and somatic mutagenesis in cancer cells VV Khrustalev, TA Khrustaleva, VV Poboinev, KV Yurchenko Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis 815, 1-9 , 2019 2019 Citations: 7
Filamentous versus Spherical Morphology: A Case Study of the Recombinant A/WSN/33 (H1N1) Virus LV Kordyukova, RR Mintaev, AA Rtishchev, MS Kunda, NN Ryzhova, ... Microscopy and Microanalysis 26 (2), 297-309 , 2020 2020 Citations: 4
Khrustalevа ТА Features of amino acid composition of beta-strands in proteins of different structural classes VV Khrustalev, VV Poboinev Fundamental’naya nauka v sovremennoi meditsine: materialy satellitnoi … , 2017 2017 Citations: 4
Особенности аминокислотного состава бета-тяжей в белках различных структурных классов ВВ Хрусталёв, ВВ Побойнев, ТА Хрусталёва Фундаментальная наука в современной медицине 2017, 331-336 , 2017 2017 Citations: 4
Peptide models of the cytoplasmic tail of influenza A/H1N1 virus hemagglutinin expand understanding its pH-dependent modes of interaction with matrix protein M1 VV Poboinev, VV Khrustalev, AA Akunevich, NV Shalygo, AN Stojarov, ... The Protein Journal 42 (4), 288-304 , 2023 2023 Citations: 3
Equilibrium between dimeric and monomeric forms of human epidermal growth factor is shifted towards dimers in a solution AA Akunevich, VV Khrustalev, TA Khrustaleva, VV Poboinev, NV Shalygo, ... The Protein Journal 41 (2), 245-259 , 2022 2022 Citations: 3
Инновационные технологии при изучении медицинской химии ТВ Прохорова, ТВ Латушко, ВВ Побойнев ББК 28.073 я431 С 23, 209 , 2022 2022 Citations: 3
Cobalt (II) cation binding by proteins. Metallomics 11, 1743–1752 VV Khrustalev, TA Khrustaleva, VV Poboinev, CI Karchevskaya, ... 2019 Citations: 3
Stability of alpha-helical and beta-structural blocks in proteins of four structural classes VV Poboinev, VV Khrustalev, TA Khrustaleva, AN Stozharov Proceedings of the National Academy of Sciences of Belarus, Biological … , 2018 2018 Citations: 3
Structural shifts of the Parvovirus B19 capsid receptor-binding domain: a peptide study VV Khrustalev, AN Stojarov, AA Akunevich, OE Baranov, AV Popinako, ... Protein and Peptide Letters 31 (2), 128-140 , 2024 2024 Citations: 2
Стабильность альфа-спиральных и бета-структурных блоков в белках четырех структурных классов ВВ Побойнев, ВВ Хрусталёв, ТА Хрусталёва, АН Стожаров Известия Национальной академии наук Беларуси. Серия биологических наук 63 (4 … , 2018 2018 Citations: 2
Особенности аминокислотного состава альфа-спиральных участков в полипептидных цепях белков различных структурных классов ВВ Побойнев, ВВ Хрусталев, ТА Хрусталева Известия Национальной академии наук Беларуси. Серия биологических наук, 58-66 , 2017 2017 Citations: 2
