mimmunМедицинская иммунологияMedical Immunology (Russia)1563-06252313-741XSPb RAACI10.15789/1563-0625-IOS-2455mimmun-2455Research ArticleОРИГИНАЛЬНЫЕ СТАТЬИORIGINAL ARTICLESИдентификация мутаций поверхностного гликопротеина SARS-CoV-2 в штаммах, изолированных в ИракеIdentification of Surface Glycoprotein Mutations of SARS-CoV-2 in Isolated Strains from Iraqhttps://orcid.org/0000-0001-8988-5957ДавудАли А.DawoodAli A.

Давуд Али А.— кандидат микробиологических наук, лектор, кафедра анатомии Медицинского колледжа Мосульского университета.

Аль-Джамеа, 1, Мосул.

Тел.: 00964 (770)-176-8002.

Ali Adel Dawood - PhD (Microbiology), Lector, Department of Anatomy, College of Medicine, University of Mosul.

Al-Jameaa, st. 1, Mosul, Iraq.

Phone: 00964 (770)-176-8002.

aad@uomosul.edu.iqhttps://orcid.org/0000-0002-1144-7444ЯсимБ. И.JasimB. I.

Ясим Бассам Исмаил.

Мосул.

Bassam Ismael Jasim.

Mosul.

bassam.jasim@uoninevah.edu.iqРияд-аль-ДжалилиО.Riadh Al-JalilyO.

Рияд-аль-Джалили Омар.

Мосул.

Omar Riadh Al-Jalily.

Mosul.

orh@uomosul.edu.iqМосульсий университетИракUniversity of MosulIraqНиневийский университетИракNinevah UniversityIraq202213072022244729740Copyright © Давуд А.А., Ясим Б.И., Рияд-аль-Джалили О., 20222022Давуд А.А., Ясим Б.И., Рияд-аль-Джалили О.Dawood A.A., Jasim B.I., Riadh Al-Jalily O.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.mimmun.ru/mimmun/article/view/2455

Глобальная пандемия коронавирусной инфекции стала длительной кризисной ситуацией для общества, экономики и здравоохранения, которая продолжается и сейчас. Спайк-гликопротеин вируса SARS-CoV-2 является одним из первичных компонентов вирулентности, тканевого тропизма и объектов носительства. Целью работы было определение мутаций S-белка в изолятах от больных COVID-19 в Ираке. Методы: полногеномные последовательности линий вируса в Ираке получали из базы GISAID. Используя статистики сатурационного мутагенеза и другие методики биинформатики, мы изучили 20 последовательностей изолятов SARS-CoV-2 с миссенс-мутацией данного белка, выявленных в Ираке и выбранных из базы данных NCBI. Результаты: во всех линиях вируса, при сравнении с диким типом, были выявлены следующие мутации: L452R, A522V, E583D and D614G. Число мутаций этих линий было различным, в зависимости от места сбора образцов. Мутация D614G была обнаружена в 19 линиях. Одна из линий имела 3 мутации, тогда как другая относились к дикому типу вируса. Структура мутантного белка существенно изменяется из-за энергетических взаимодействий атомов в зоне стыковки, что влияет на стабильность белка. Выводы: стабильность S-белка может изменяться в зависимости от места мутации. Стыковка молекулы RBD-ACE2 нарушается по-разному при заменах аминокислот L452R and A522V.

Background: The global pandemic of coronavirus disease is a societal, economic, and publichealth crisis that is still underway. The spike glycoprotein of SARS-CoV-2 is one of the primary ingredients for virulence, tissue tropism, and host areas. Aim: This study aimed to determine mutations in the S protein of the Iraqi COVID-19 isolates.

Full genome sequences of Iraqi strains were obtained from GISAID. Using statistical saturation mutagenesis and other informatics methods, we investigated 20 sequences of SARS-CoV-2 S protein missense mutation isolates in Iraq selected from NCBI.

The following mutations were detected for all the strains under study compared to the wild type: L452R, A522V, E583D and D614G. The number of mutations in the strains was different depending on the location of the state from which the sample was collected The D614G mutation was found in 19 strains. One strain had three mutations, while the other was a wild form strain. The structure of the mutant protein changes dramatically, as does the energy of the atoms concerning the docking position, affecting the protein's stability.

The mutation sites would improve the S protein's stability. Molecular docking of RBD-ACE2 is affected differently by residues L452R and A522V.

SARS-CoV-2спайк-протеинмутацииповерхностныеACE2RBDSARS-CoV-2SpikeMutationSurfaceACE2RBDThe authors express their gratitude to the medical colleges of Nineveh and Mosul for recording their work.ReferencesBestle D., Heindl M.R., Limburg H., Pilgram O., Moulton H., Stein D.A., Hardes K., Eickmann M., Dolnik O., Rohde C., Klenk H.-D., Garten W, Steinmetzer T., Bottcher-Friebertshauser E. TMPRSS2 and furin are both essential for proteolytic activation and spread of SARS-CoV-2 in human airway epithelial cells and provide promising drug targets. Life Sci. Alliance, 2020, Vol. 3, e202000786. doi: 10.26508/lsa.202000786.Lan J, Ge J, Shan S, Zhou H, Fan S, and Zhang Q. et al. Structure of the SARS-CoV-2 spike receptor-binding domain bound to the ACE2 receptor. Nature. 2020; 581, 215-231. DOI: 10.1038/s41586-020-2180-5. PMID: 32225176.Bosch B.J., van der Zee R., de Haan C.A., Rottier P.J. The coronavirus spike protein is a class I virus fusion protein: structural and functional characterization of the fusion core complex. J. Virol., 2003, Vol. 77, no. 16, pp. 88018811.Dawood A. Identification of CTL and B-cell epitopes in the Nucleocapsid Phosphoprotein of COVID-19 using Immunoinformatics. Microbiol J. 2021; 83(1): 78-86. DOI: 10.15407/microbiolj83.01.078.Calcagnile M., Forgez P., Lannelli A., Bucci C., Alifano M., Alifano P. Molecular docking simulation reveals ACE2 polymorphisms that may increase the affinity of ACE2 with the SARS-CoV-2 Spike protein. Biochimie, 2021, Vol. 180, pp. 143-148.Dawood A. Glycosylation, ligand binding sites and antigenic variations between membrane glycoprotein of COVID-19 and related coronaviruses. Vacunas. 2021; 22(1): 1-9. DOI: 10.1016/j.vacun.2020.09.005. PMID: 33041736.Chan K., Dorosky D., Shamra P., Abbasi S., Dye J., Kranz D., Herbert A.S. Procko E. Engineering human ACE2 to optimize binding to the spike protein of SARS coronavirus 2. Science, 2020, Vol. 369, no. 6508, pp. 1261-1265.Shang J, Ye G, Shi K, Wan Y, Luo C, and Aihara H. et al. Structural basis of receptor ecognition by SARS-CoV-2. Nature. 2020; 581, 221-239. DOI: 10.1038/s41586-020-2179- y. PMID: 32225175.Dawood A. Glycosylation, ligand binding sites and antigenic variations between membrane glycoprotein of COVID-19 and related coronaviruses. Vacunas, 2021, Vol. 22, no. 1, pp. 1-9.Zhang L, Jackson C, Mou H, Ojha A, Quinlan B, and Ranarajan E. et al. SARS-CoV-2 spike- protein D614G mutation increases virion spike density and infectivity. Nature Comm. 2020; 11:6013. DOI: 10.1038/s41467-020-19808-4.Dawood A. Identification of CTL and B-cell epitopes in the Nucleocapsid Phosphoprotein of COVID-19 using Immunoinformatics. Microbiol. J., 2021, Vol. 83, no. 1, pp. 78-86.Ou X, Liu Y, Lei X, Li P, Mi D, and Ren L. et al. Characterization of spike glycoprotein of SARS-CoV-2 on virus entry and its immune cross-reactivity with SARS-CoV. Nature Comm. 2020; 11:1620. DOI: 10.1038/s41467-020-15562-9. PMID: 32221306.Dawood A. Mutated COVID-19, May Foretells Mankind in a Great Risk in The Future. New Microbes New Infect., 2020, Vol. 35, 100673. doi: 10.1016/j.nmni.2020.100673.Dawood A, Altobje M. Inhibition of N-linked Glycosylation by Tunicamycin May Contribute to The Treatment of SARS-CoV-2. Microbiol Path. 2020; 149:104586. DOI: 10.1016/j.micpath.2020.104586.Dawood A., Altobje M. Inhibition of N-linked Glycosylation by Tunicamycin May Contribute to The Treatment of SARS-CoV-2. Microbiol. Path., 2020, Vol. 149, 104586. doi: 10.1016/j.micpath.2020.104586.Duan L, Zheng Q, Zhang H, Nlu Y, and Wang H. The SARS-CoV-2 Spike Glycoprotein Biosynthesis, Structure, Function, and Antigenicity: Implications for the Design of Spike- Based Vaccine Immunogens. Front. Immunol. 2020; 11, 1-12. DOI: 10.3389/fimmu.2020.576622.Dawood A., Altobje M., Alnori H. Compatibility of the ligand binding sites in the spike glycoprotein of COVID-19 with those in the aminopeptidase and the caveolins 1, 2 proteins. Res. J. Pharm. Tech., 2021, Vol. 14, no. 9, pp. 4760-4766. Wu F., Zhao S., Yu B., Chen Y.M, Wang W., Song Z.G., Hu Y., Tao Z.-W., Tian J.-H., Pei Y.-Y., Yuan M.-L., Zhang Y.-L., Dai F.-H., Liu Y., Wang Q.-M., Zheng J.-J., Xu L., Holmes E.C., Zhang Y.-Z. A new coronavirus associated with human respiratory disease in China. Nature, 2020, Vol. 579, pp. 265-269.Othman H, Bouslama Z, Brandenburg JT, Rocha J, Hamdi Y, Ghedira K. et al. Interaction of the spike protein RBD from SARS-CoV-2 with ACE2: Similarity with SARS-CoV, hot- spot analysis and effect of the receptor polymorphism. Biochem. Biophys. Res. Comm. 2020; 527,702-710. DOI: 10.1016/j.bbrc.2020.05.028. PMID: 32410735.Dawood A., Altobje M., Alrassam Z. Molecular Docking of SARS-CoV-2 nucleocapsid protein with angiotensin-converting enzyme II. Mikrobio Zhu., 2021, Vol. 83, no. 2, pp. 82-92.Durmaz B, Abdulmajed O, and Durmaz R. Mutations Observed in the SARS-CoV-2 Spike Glycoprotein and Their Effects in the Interaction of Virus with ACE-2 Receptor. Medeni Med J. 2020; 35, 235-260. DOI: 10.5222/MMJ.2020.98048. PMID: 33110678.Dieterle M.E., Haslwanter D., Bortz III R.H., Wirchnianski A.S., Lasso G., Vergnolle O., Abbasi S.A., Fels J.M., Laudermilch E., Florez C., Mengotto A., Kimmel D., Malonis R.J., Georgiev G., Quiroz J., Barnhill J., Pirofski L.-A., Daily J.P., Dye J.M., Lai J.R., Herbert A.S., Chandran K., Jangra R.K. A replication-competent vesicular stomatitis virus for studies of SARS-CoV-2 spike-mediated cell entry and its inhibition. Cell Host Microbe, 2020, Vol. 28, no. 3, pp. 486-496.e6.Li Q, Wu J, Nie J, Li X, Huang W, and Wang Y. The Impact of Mutations in SARS-CoV-Duan L., Zheng Q., Zhang H., Nlu Y., Wang H. The SARS-CoV-2 spike glycoprotein biosynthesis, structure, function, and antigenicity: implications for the design of spike-based vaccine immunogens. Front. Immunol. 2020, Vol. 11, 576622. doi: 10.3389/fimmu.2020.576622.Spike on Viral Infectivity and Antigenicity. Cell. 2020; 182, 1248-1294. DOI: 10.1016/j.cell.2020.07.012. PMID: 32730807.Durmaz B., Abdulmajed O., Durmaz R. Mutations observed in the SARS-CoV-2 spike glycoprotein and their effects in the interaction of virus with ACE-2 receptor. Medeni Med. J., 2020, Vol. 35, no. 3, pp. 253-260.Hoffmann M., Kleine-Weber H., Schroeder S., Kruger N., Herrler T., Erichsen S., Schiergens T.S., Herrler G., Wu N.-H., Nitsche A., Muller M.A. Drosten C., Pohlmann S. SARS- CoV-2 cell entry depends on ACE2 and TMPRSS2 and is blocked by a clinically proven protease inhibitor. Cell, 2020, Vol. 181, no. 2, pp. 271-280.e8.Dawood A. Mutated COVID-19, May Foretells Mankind in a Great Risk in The Future. N Mic N Inf. 2020; 35. DOI: 10.1016/j.nmni.2020.100673. PMID: 32292587Iwanaga N., Cooper L., Rong L., Beddingfield B., Crabtree J., Tripp R.A., Qin X., Kolls J.K. Novel ACE2-IgG1 fusions with increased activity against SARS-CoV-2. bioRxiv, 2020. doi: 10.1101/2020.06.15.152157.Othman H, Bouslama Z, Brandenburg J, Rocha J, Hamdi Y, and Ghedira K. et al. In silico study of the spike protein from SARS-CoV-2 interaction with ACE2: similarity with SARS- CoV, hot-spot analysis and effect of the receptor polymorphism. bioRxiv. 2020; 339781420. DOI: 10.1101/2020.03.04.976027. PMID: 32410735.Lan J., Ge J., Shan S., Zhou H., Fan S., Zhang Q., Shi X., Wang Q., Zhang L., Wang X. Structure of the SARS-CoV-2 spike receptor-binding domain bound to the ACE2 receptor. Nature, 2020, Vol. 581, no. 7807, pp. 215-231.Chan K, Dorosky D, Shamra P, Abbasi S, Dye J, and Kranz D et al. Engineering human ACE2 to optimize binding to the spike protein of SARS coronavirus 2. Ortho Biolo. 2020; 369, 1261-1265. PMID: 32753553.Lan J., Ge J., Yu J., Shan S., Zhou H., Fan S., Zhang Q., Shi X., Wang Q., Zhang L., Wang X. Structure of the SARS-CoV-2 spike receptor-binding domain bound to the ACE2 receptor. Nature, 2020, Vol. 581, no. 7807, pp. 215-220.Calcagnile M, Forgez P, Lannelli A, Bucci C, Alifano M, and Alifano P. Molecular docking simulation reveals ACE2 polymorphisms that may increase the affinity of ACE2 with the SARS-CoV-2 Spike protein. Biochimie. 2021; 180, 143-148. DOI: 10.1016/j.biochi.2020.11.004. PMID: 33181224.Lei C., Qian K., Li T., Zhang S., Fu W., Ding M., Hu S. Neutralization of SARSCoV-2 spike pseudotyped virus by recombinant ACE2-Ig. Nat. Commun., 2020, Vol. 11, no. 1, 2070. doi: 10.1038/s41467-020-16048-4.Bestle D, Heindl MR, Limburg H, Pilgram O, Moulton H, Stein DA, et al. TMPRSS2 and furin are both essential for proteolytic activation and spread of SARS-CoV-2 in human airway epithelial cells and provide promising drug targets. Life Sci Alliance. 2020; 3:e202000786. DOI: 10.26508/lsa.202000786. PMID: 32703818.Letko M., Marzi A., Munster V. Functional assessment of cell entry and receptor usage for SARS-CoV-2 and other lineage B betacoronaviruses. Nat. Microbiol., 2020, Vol. 5, pp. 562-569.Lan J, Ge J, Yu J, Shan S, Zhou H, Fan S, et al. Structure of the SARS-CoV-2 spike receptor- binding domain bound to the ACE2 receptor. Nature. 2020; 581:215–20. DOI: 10.1038/s41586-020-2180-5. PMID: 32225176.Liang Y., Wang M.-L., Chien C.-S., Yarmishyn A.A., Yang Y.-P., Lai W.-Y., LuoY .-H., Lin Y.-T., Chen Y.-J., Chang P.-C., Chiou S.-H. Highlight of immune pathogenic response and hematopathologic effect in SARS-CoV, MERS-CoV, and SARS-Cov-2 infection. Front. Immunol., 2020, Vol. 11, 1022. doi: 10.3389/fimmu.2020.01022.Dawood A, Altobje M, and Alnori H. Compatibility of the Ligand Binding Sites in the Spike Glycoprotein of COVID-19 with those in the Aminopeptidase and the Caveolins 1, 2 Proteins. Res J Pharm Tech. 2021; 14(9): 4760-4766. doi:10.52711/0974- 360X.2021.00828.Wu F, Zhao S, Yu B, Chen YM, Wang W, Song ZG, et al. A new coronavirus associated with human respiratory disease in China. Nature. 2020; 579:265–9. DOI: 10.1038/s41586-020-2008-3.Li Q., Wu J., Nie J., Zhang L., Hao H., Liu S., Zhao C., Zhang Q., Liu H., Nie L., Qin H., Wang M., Lu Q., Li X., Sun Q., Liu J., Zhang L., Li X., Huang W., Wang Y. The impact of mutations in SARS-CoV-2 spike on viral infectivity and antigenicity. Cell, 2020, Vol. 182, no. 5, pp. 1248-1294.Liang Y, Wang M-L, Chien C-S, Yarmishyn AA, Yang Y-P, Lai W-Y, et al. Highlight of Immune Pathogenic Response and Hematopathologic Effect in SARS-CoV, MERS-CoV, and SARS-Cov-2 Infection. Front Immunolo. 2020; 11:1022. DOI: 10.3389/fimmu.2020.01022. PMID: 32574260.Malik Y.A. Properties of Coronavirus and SARS-CoV-2. Malays. J. Pathol., 2020, Vol. 42, no. 1, pp. 3-11.Tang T, Bidon M, Jaimes JA, Whittaker GR, and Daniel S. Coronavirus membrane fusion mechanism offers apotential target for antiviral development. . 2020; 178:104792. DOI: 10.1016/j.antiviral. 2020.104792. PMID: 32272173.Monteil V., Kwon H., Prado P., Hagelkruys A., Wimmer R.A., Stahl M., Leopoldi A., Garreta E., Hurtado Del Pozo C., Prosper F., Romero J.P , Wirnsberger G., Zhang H., Slutsky A.S., Conder R., Montserrat N., Mirazimi A., Penninger J.M. Inhibition of SARS-CoV-2 infections in engineered human tissues using clinical-grade soluble human ACE2. Cell, 2020, Vol. 181, no. 4, pp. 905-913.e7.Hoffmann M, Kleine-Weber H, Schroeder S, Kruger N, Herrler T, Erichsen S, et al. SARS- CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a Clinically Proven Protease Inhibitor. Cell. 2020; 181:271–80.e8. DOI: 10.1016/j.cell.2020.02.052. PMID: 32142651.Othman H., Bouslama Z., Brandenburg J.T., Rocha J., Hamdi Y., Ghedira K., Srairi-Abid N., Hazelhurst S. Interaction of the spike protein RBD from SARS-CoV-2 with ACE2: Similarity with SARS-CoV, hot- spot analysis and effect of the receptor polymorphism. Biochem. Biophys. Res. Comm., 2020, Vol. 527, no. 3, pp. 702-710.Letko M, Marzi A, Munster V. Functional assessment of cell entry and receptor usage for SARS-CoV-2 and other lineage B betacoronaviruses. Nat. Microbiol. 2020; 5:562–9. DOI: 10.1038/s41564-020-0688-y. PMID: 32094589.Othman H., Bouslama Z., Brandenburg J.T., Rocha J., Hamdi Y., Ghedira K., Srairi-Abid N., Hazelhurst S. In silico study of the spike protein from SARS-CoV-2 interaction with ACE2: similarity with SARS- CoV, hot-spot analysis and effect of the receptor polymorphism. BioRxiv, 2020, 339781420. doi: 10.1101/2020.03.04.976027.Shang J, Ye G, Shi K, Wan Y, Luo C, Aihara H, et al. Structural basis of receptor recognition by SARS-CoV-2. Nature. 2020; 581:221–4. DOI: 10.1038/s41586-020-2179-y. PMID: 32225175.Ou X., Liu Y., Lei X., Li P., Mi D., Ren L., Guo L., Guo R., Chen T., Hu J., Xiang Z., Mu Z., Chen X., Chen J., Hu K., Jin Q., Wang J., Qian Z. Characterization of spike glycoprotein of SARS-CoV-2 on virus entry and its immune cross-reactivity with SARS-CoV. Nature Comm., 2020, Vol. 11, no. 1, 1620. doi: 10.1038/s41467-020-15562-9.Schmidt F., Weisblum Y., Muecksch F., Hoffmann H.-H., Michailidis E., Lorenzi J.C., Mendoza P., Rutkowska M., Bednarski E., Gaebler C., Agudelo M., Cho A., Wang Z., Gazumyan A., Cipolla M., Caskey M., Robbiani D.F., Nussenzweig M.C., Rice C.M., Hatziioannou T., Bieniasz P.D. Measuring SARS-CoV-2 neutralizing antibody activity using pseudotyped and chimeric viruses. J. Exp. Med., 2020, Vol. 217, no. 11, e20201181. doi: 10.1084/jem.20201181.Lan J, Ge J, Yu J, Shan S, Zhou H, Fan S, et al. Structure of the SARS-CoV-2 spike receptor- binding domain bound to the ACE2 receptor. Nature.2020; 581:215–20. doi: 10.1038/s41586-020-2180-5. PMID: 32225176.Shang J., Ye G., Shi K., Wan Y., Luo C., Aihara H., Geng Q., Auerbach A., Li F. Structural basis of receptor ecognition by SARS-CoV-2. Nature, 2020, Vol. 581, no. 7807, pp. 221-239.Wang Q, Zhang Y, Wu L, Niu S, Song C, Zhang Z, et al. Structural and Functional Basis of SARS-CoV-2 Entry by Using Human ACE2. Cell. 2020; 181:894–904 e9. DOI: 10.1016/j.cell.2020.03.045. PMID: 32275855.Tai W., He L., Zhang X., Pu J., Voronin D., Jiang S., Zhou Y., Du L. Characterization of the receptor-binding domain (RBD) of 2019 novel coronavirus: implication for development of RBD protein as a viral attachment inhibitor and vaccine. Cell. Mole. Immunol., 2020, Vol. 17, no. 6, pp. 613-620.Yan R, Zhang Y, Li Y, Xia L, Guo Y, Zhou Q. Structural basis for the recognition of SARS- CoV-2 by full-length human ACE2. Science. 2020; 367:1444–8. DOI: 10.1126/science.abb2762. PMID: 32132184.Tang T., Bidon M., Jaimes J.A., Whittaker G.R., Daniel S. Coronavirus membrane fusion mechanism offers apotential target for antiviral development. Antiviral Res., 2020, Vol. 178, 104792. doi: 10.1016/j.antiviral.2020.104792.Monteil V, Kwon H, Prado P, Hagelkruys A, Wimmer RA, Stahl M, et al. Inhibition of SARS-CoV-2 Infections in Engineered Human Tissues Using Clinical-Grade Soluble Human ACE2. Cell. 2020; 181:905–13 e7. DOI: 10.1016/j.cell.2020.04.004. PMID: 32333836.Teng S., Sobitan A., Rhoades R., Liu D., Tang Q. Systemic effects of missense mutations on SARS-CoV-2 spike glycoprotein stability and receptor-binding affinity. Brief Bioinf., 2021, Vol. 22, no. 2, pp. 1239-1253.Lei C, Qian K, Li T, Zhang S, Fu W, Ding M, et al. Neutralization of SARSCoV-2 spike pseudotyped virus by recombinant ACE2-Ig. Nat. Commun. 2020; 11:2070. DOI: 10.1038/s41467-020-16048-4. PMID: 32332765.Ujike M., Huang C., Shirato K., Makino S., Taguchi F. The contribution of the cytoplasmic retrieval signal of severe acute respiratory syndrome coronavirus to intracellular accumulation of S proteins and incorporation of S protein into viruslike particles. J. Gen. Virol., 2016, Vol. 97, no. 8, pp. 1853-1864.Iwanaga N, Cooper L, Rong L, Beddingfield B, Crabtree J, Tripp RA, et al. Novel ACE2- IgG1 fusions with increased activity against SARS-CoV-2. bioRxiv. 2020. DOI: 10.1101/2020.06.15.152157. PMID: 32587964.Wang Q., Zhang Y., Wu L., Niu S., Song C., Zhang Z., Lu G., Qiao C., Hu Y., Yuen K.-Y. , Wang Q., Zhou H., Yan J., Qi J. Structural and Functional Basis of SARS-CoV-2 Entry by Using Human ACE2. Cell, 2020, Vol. 181, no. 4, pp. 894-904.e9.Zhang G, Pomplun S, Loftis AR, Loas A, Pentelute B. Thefirst-in-class peptide binder to the SARS-CoV-2 spike protein. bioRxiv. 2020. DOI: 10.1101/2020.03.19.999318.Wrapp D., Wang N., Corbett K.S., Goldsmith J.A., Hsieh C.L., Abiona O., Graham B.S. , McLellan J.S. CryoEM structure of the 2019-nCoV spike in the prefusion conformation. Science, 2020, Vol. 367, no. 6483, pp. 1260-1263.Ujike M, Huang C, Shirato K, Makino S, Taguchi F. The contribution of the cytoplasmic retrieval signal of severe acute respiratory syndrome coronavirus to intracellular accumulation of S proteins and incorporation of S protein into viruslike particles. J. Gen. Virol. 2016; 97:1853–64. DOI: 10.1099/jgv.0.000494. PMID: 27145752.Wu F., Zhao S., Yu B., Chen Y.M., Wang W., Song Z.G, A new coronavirus associated with human respiratory disease in China. Nature, 2020, Vol. 579, pp. 265-269.Malik YA. Properties of Coronavirus and SARS-CoV-2. Malays. J. Pathol. 2020; 42:3–11. 32. PMID: 32342926. PMID: 32342926.Yan R., Zhang Y., Li Y., Xia L., Guo Y., Zhou Q. Structural basis for the recognition of SARS-CoV-2 by fulllength human ACE2. Science, 2020, Vol. 367, no. 6485, pp. 1444-1448.Dieterle ME, Haslwanter D, Bortz III RH, Wirchnianski AS, Lasso G, Vergnolle O, et al. A replication-competent vesicular stomatitis virus for studies of SARS-CoV-2 spike-mediated cell entry and its inhibition. Cell Host Microbe. 2020; 28:486–96.e6. DOI: 0.1016/j.chom.2020.06.020. PMID: 32738193.Zhang G., Pomplun S., Loftis A.R., Loas A., Pentelute B. Thefirst-in-class peptide binder to the SARS-CoV-2 spike protein. bioRxiv. 2020. doi: 10.1101/2020.03.19.999318.Schmidt F, Weisblum Y, Muecksch F, Hoffmann H-H, Michailidis E, Lorenzi JC, et al. Measuring SARS-CoV-2 neutralizing antibody activity using pseudotyped and chimeric viruses. J. Exp. Med. 2020; 217:e20201181. DOI: 10.1084/jem.20201181. PMID: 32692348.Zhang L., Jackson C., Mou H., Ojha A., Quinlan B., Ranarajan E., Izard T., Farzan M., Choe H. SARS-CoV-2 spike- protein D614G mutation increases virion spike density and infectivity. Nature Comm., 2020, Vol. 11, 6013. doi: 10.1038/s41467-020-19808-4.Wrapp D, Wang N, Corbett KS, Goldsmith JA, Hsieh CL, Abiona O, et al. Cryo-EM structure of the 2019-nCoV spike in the prefusion conformation. Science. 2020; 367:1260– 3. DOI: 10.1126/science.abb2507. PMID: 32075877.Tai W, He L, Zhang X, Pu J, Voronin D, and Jiang S. et al. Characterization of the receptor- binding domain (RBD) of 2019 novel coronavirus: implication for development of RBD protein as a viral attachment inhibitor and vaccine. Cell. Mole. Immunol. 2020; 17:613–620. DOI: 10.1038/s41423-020-0400-4.Tai W, He L, Zhang X, Pu J, Voronin D, and Jiang S. et al. Characterization of the receptor- binding domain (RBD) of 2019 novel coronavirus: implication for development of RBD protein as a viral attachment inhibitor and vaccine. Cell. Mole. Immunol. 2020; 17:613–620. DOI: 10.1038/s41423-020-0400-4.Bosch BJ, van der Zee R, de Haan CA, Rottier PJ. The coronavirus spike protein is a class I virus fusion protein: structural and functional characterization of the fusion core complex. J. Virol. 2003; 77:8801–11. DOI: 10.1128/jvi.77.16.8801-8811.2003.Bosch BJ, van der Zee R, de Haan CA, Rottier PJ. The coronavirus spike protein is a class I virus fusion protein: structural and functional characterization of the fusion core complex. J. Virol. 2003; 77:8801–11. DOI: 10.1128/jvi.77.16.8801-8811.2003.Dawood A., Altobje M., Alrassam Z. Molecular Docking of SARS-CoV-2 Nucleocapsid Protein with Angiotensin-Converting Enzyme II. Mikrobio Zhu. 2021; 83(2):82-92. DOI: 10.15407/microbiolj83.02.082.Dawood A., Altobje M., Alrassam Z. Molecular Docking of SARS-CoV-2 Nucleocapsid Protein with Angiotensin-Converting Enzyme II. Mikrobio Zhu. 2021; 83(2):82-92. DOI: 10.15407/microbiolj83.02.082.Teng S, Sobitan A, Rhoades R, Liu D, and Tang Q. Systemic effects of missense mutations on SARS-CoV-2 spike glycoprotein stability and receptor-binding affinity. Brief. Bioinf. 2021; 22(2), 1239-1253. DOI: 10.1093/bib/bbaa233. PMID: 33006605.Teng S, Sobitan A, Rhoades R, Liu D, and Tang Q. Systemic effects of missense mutations on SARS-CoV-2 spike glycoprotein stability and receptor-binding affinity. Brief. Bioinf. 2021; 22(2), 1239-1253. DOI: 10.1093/bib/bbaa233. PMID: 33006605.

The authors declare that there are no conflicts of interest present.