Development, validation and analysis of a human profurin 3D model using comparative modeling and molecular dynamics simulations

References

• Abraham, M. J., Murtola, T., Schulz, R., Páll, S., Smith, J. C., Hess, B., & Lindahl, E. (2015). Gromacs: High performance molecular simulations through multi-level parallelism from laptops to supercomputers. SoftwareX, 1–2, 19–25. https://doi.org/10.1016/j.softx.2015.06.001
   Google Scholar
• Altschul, S. F., Gish, W., Miller, W., Myers, E. W., & Lipman, D. J. (1990). Basic local alignment search tool. Journal of Molecular Biology, 215(3), 403–410. https://doi.org/10.1016/S0022-2836(05)80360-2
   PubMed Web of Science ®Google Scholar
• Amadei, A., Linssen, A. B., & Berendsen, H. J. (1993). Essential dynamics of proteins. Proteins: Structure, Function, and Bioinformatics, 17(4), 412–425. https://doi.org/10.1002/prot.340170408
   PubMed Web of Science ®Google Scholar
• Berendsen, H. J., Postma, J. P., & van Gunsteren, W. F. (1981). Interaction models for water in relation to protein hydration. In: Intermolecular forces (pp. 331–342). Springer.
   Google Scholar
• Berman, H. M., Westbrook, J., Feng, Z., Gilliland, G., Bhat, T. N., Weissig, H., Shindyalov, I. N., & Bourne, P. E. (2000). The protein data bank. Nucleic Acids Research, 28(1), 235–242. https://doi.org/10.1093/nar/28.1.235
   PubMed Web of Science ®Google Scholar
• Best, R. B., Zhu, X., Shim, J., Lopes, P. E. M., Mittal, J., Feig, M., & MacKerell, A. D. (2012). Optimization of the additive charmm all-atom protein force field targeting improved sampling of the backbone ϕ, ψ and side-chain χ1 and χ2 dihedral angles. Journal of Chemical Theory and Computation, 8(9), 3257–3273. https://doi.org/10.1021/ct300400x
   PubMed Web of Science ®Google Scholar
• Bhattacharjya, S., Xu, P., Wang, P., Osborne, M. J., & Ni, F. (2007). Conformational analyses of a partially-folded bioactive prodomain of human furin. Biopolymers: Original Research on Biomolecules, 86(4), 329–344. https://doi.org/10.1002/bip.20748
   PubMed Web of Science ®Google Scholar
• Braun, E., & Sauter, D. (2019). Furin-mediated protein processing in infectious diseases and cancer. Clinical & Translational Immunology, 8(8), e1073. https://doi.org/10.1002/cti2.1073
   PubMed Web of Science ®Google Scholar
• Bussi, G., Donadio, D., & Parrinello, M. (2007). Canonical sampling through velocity rescaling. The Journal of Chemical Physics, 126(1), 014101. https://doi.org/10.1063/1.2408420
   PubMed Web of Science ®Google Scholar
• Case, D. A. (2022). Amber 2022. https://ambermd.org/doc12/Amber22.pdf#page=33
   Google Scholar
• Chen, V. B., Arendall, W. B., Headd, J. J., Keedy, D. A., Immormino, R. M., Kapral, G. J., Murray, L. W., Richardson, J. S., & Richardson, D. C. (2010). Molprobity: All-atom structure validation for macromolecular crystallography. Acta Crystallographica. Section D, Biological Crystallography, 66(Pt 1), 12–21. https://doi.org/10.1107/S0907444909042073
   PubMed Web of Science ®Google Scholar
• Cino, E. A., Choy, W. Y., & Karttunen, M. (2012). Comparison of secondary structure formation using 10 different force fields in microsecond molecular dynamics simulations. Journal of Chemical Theory and Computation, 8(8), 2725–2740. https://doi.org/10.1021/ct300323g
   PubMed Web of Science ®Google Scholar
• Colovos, C., & Yeates, T. O. (1993). Verification of protein structures: Patterns of nonbonded atomic interactions. Protein Science: A Publication of the Protein Society, 2(9), 1511–1519. https://doi.org/10.1002/pro.5560020916
   PubMed Web of Science ®Google Scholar
• da Silva, T. U., Pougy, K. D. C., Albuquerque, M. G., Lima, C. H. D. S., & Machado, S. D. P. (2023). Molecular dynamics simulations of aqueous systems of inhibitor candidates for adenosine-5’-phosphosufate reductase. Journal of Biomolecular Structure and Dynamics, 41(6), 2466–2477. https://doi.org/10.1080/07391102.2022.2033137
   PubMed Web of Science ®Google Scholar
• Darden, T., York, D., & Pedersen, L. (1993). Particle mesh ewald: An n log (n) method for ewald sums in large systems. The Journal of Chemical Physics, 98(12), 10089–10092. https://doi.org/10.1063/1.464397
   Web of Science ®Google Scholar
• David, C. C., & Jacobs, D. J. (2014). Principal component analysis: a method for determining the essential dynamics of proteins. In Protein dynamics (pp. 193–226). Springer.
   Google Scholar
• de Souza, A. S., Pacheco, B. D. C., Pinheiro, S., Muri, E. M. F., Dias, L. R. S., Lima, C. H. S., Garrett, R., de Moraes, M. B. M., de Souza, B. E. G., & Puzer, L. (2019). 3-acyltetramic acids as a novel class of inhibitors for human kallikreins 5 and 7. Bioorganic & Medicinal Chemistry Letters, 29(9), 1094–1098. https://doi.org/10.1016/j.bmcl.2019.02.031
   PubMed Web of Science ®Google Scholar
• Dillon, S. L., Williamson, D. M., Elferich, J., Radler, D., Joshi, R., Thomas, G., & Shinde, U. (2012). Propeptides are sufficient to regulate organelle-specific pH-dependent activation of furin and proprotein convertase 1/3. Journal of Molecular Biology, 423(1), 47–62. https://doi.org/10.1016/j.jmb.2012.06.023
   PubMed Web of Science ®Google Scholar
• Dong, G. Q., Fan, H., Schneidman-Duhovny, D., Webb, B., & Sali, A. (2013). Optimized atomic statistical potentials: Assessment of protein interfaces and loops. Bioinformatics (Oxford, England), 29(24), 3158–3166. https://doi.org/10.1093/bioinformatics/btt560
   PubMed Web of Science ®Google Scholar
• Duan, Y., Wu, C., Chowdhury, S., Lee, M. C., Xiong, G., Zhang, W., Yang, R., Cieplak, P., Luo, R., Lee, T., Caldwell, J., Wang, J., & Kollman, P. (2003). A point-charge force field for molecular mechanics simulations of proteins based on condensed-phase quantum mechanical calculations. Journal of Computational Chemistry, 24(16), 1999–2012. https://doi.org/10.1002/jcc.10349
   PubMed Web of Science ®Google Scholar
• Feliciangeli, S. F., Thomas, L., Scott, G. K., Subbian, E., Hung, C.-H., Molloy, S. S., Jean, F., Shinde, U., & Thomas, G. (2006). Identification of a pH sensor in the furin propeptide that regulates enzyme activation. The Journal of Biological Chemistry, 281(23), 16108–16116. https://doi.org/10.1074/jbc.M600760200
   PubMed Web of Science ®Google Scholar
• Ferrari, B. (2023). Automated-lysozyme-water-forcefields-GROMACS. https://github.com/brendaferrari/automated-lysozyme-water-forcefields-GROMACS.
   Google Scholar
• Ferrari, B. (2023). HbondAuto. https://github.com/brendaferrari/HBondAuto.
   Google Scholar
• Ferrari, B. (2023). InfleCS-analysis. https://github.com/brendaferrari/InfleCS-analysis.
   Google Scholar
• Ferrari, B. (2023). PCAauto. https://github.com/brendaferrari/PCAauto.
   Google Scholar
• Ferrari, B. (2023). PORCUPINEplot. https://github.com/brendaferrari/PORCUPINEplot.
   Google Scholar
• Fiser, A. (2010). Template-based protein structure modeling. Computational Biology, 443, 73–94.
   PubMedGoogle Scholar
• Foundation PS. (2016). Python language reference. Versão 3.6. https://www.python.org/downloads/release/python-3613/
   Google Scholar
• Foundation PS. (2022). Python language reference. Versão 3.8. https://www.python.org/downloads/release/python-3813/
   Google Scholar
• Fugère, M., Limperis, P. C., Beaulieu-Audy, V., Gagnon, F., Lavigne, P., Klarskov, K., Leduc, R., & Day, R. (2002). Inhibitory potency and specificity of subtilase-like pro-protein convertase (SPC) prodomains. The Journal of Biological Chemistry, 277(10), 7648–7656. https://doi.org/10.1074/jbc.M107467200
   PubMed Web of Science ®Google Scholar
• Gawlik, K., Shiryaev, S. A., Zhu, W., Motamedchaboki, K., Desjardins, R., Day, R., Remacle, A. G., Stec, B., & Strongin, A. Y. (2009). Autocatalytic activation of the furin zymogen requires removal of the emerging enzyme’s n-terminus from the active site. PLoS One. 4(4), e5031. https://doi.org/10.1371/journal.pone.0005031
   PubMed Web of Science ®Google Scholar
• Gowers, R. J., Linke, M., & Barnoud, J. (2016). Mdanalysis: A python package for the rapid analysis of molecular dynamics simulations. Proceedings of the 15th Python in Science Conference, 98,105. https://doi.org/10.25080/Majora-629e541a-00e
   Google Scholar
• Guvench, O., & MacKerell, A. D. (2008). Comparison of protein force fields for molecular dynamics simulations. Molecular Modeling of Proteins, 673, 63–88. [Database][Mismatch
   Google Scholar
• Hameduh, T., Haddad, Y., Adam, V., & Heger, Z. (2020). Homology modeling in the time of collective and artificial intelligence. Computational and Structural Biotechnology Journal, 18, 3494–3506. https://doi.org/10.1016/j.csbj.2020.11.007
   PubMed Web of Science ®Google Scholar
• Harris, C. R., Millman, K. J., van der Walt, S. J., Gommers, R., Virtanen, P., Cournapeau, D., Wieser, E., Taylor, J., Berg, S., Smith, N. J., Kern, R., Picus, M., Hoyer, S., van Kerkwijk, M. H., Brett, M., Haldane, A., Del Río, J. F., Wiebe, M., Peterson, P. … Oliphant, T. E. (2020). Array programming with numpy. Nature, 585(7825), 357–362. https://doi.org/10.1038/s41586-020-2649-2
   PubMed Web of Science ®Google Scholar
• Hess, B., Bekker, H., Berendsen, H. J. C., & Fraaije, J. G. E. M. (1997). Lincs: A linear constraint solver for molecular simulations. Journal of Computational Chemistry, 18(12), 1463–1472. https://doi.org/10.1002/(SICI)1096-987X(199709)18:12<1463::AID-JCC4>3.0.CO;2-H
   Web of Science ®Google Scholar
• Hornak, V., Abel, R., Okur, A., Strockbine, B., Roitberg, A., & Simmerling, C. (2006). Comparison of multiple amber force fields and development of improved protein backbone parameters. Proteins: Structure, Function, and Bioinformatics, 65(3), 712–725. https://doi.org/10.1002/prot.21123
   PubMed Web of Science ®Google Scholar
• Huang, J., & MacKerell, A. D. (2013). Charmm36 all-atom additive protein force field: Validation based on comparison to NMR data. Journal of Computational Chemistry, 34(25), 2135–2145. https://doi.org/10.1002/jcc.23354
   PubMed Web of Science ®Google Scholar
• Humphrey, W., Dalke, A., & Schulten, K. (1996). VMD: Visual molecular dynamics. Journal of Molecular Graphics, 14(1), 33–38. https://doi.org/10.1016/0263-7855(96)00018-5
   PubMedGoogle Scholar
• Hunter, J. D. (2007). Matplotlib: A 2d graphics environment. Computing in Science & Engineering, 9(3), 90–95. https://doi.org/10.1109/MCSE.2007.55
   Web of Science ®Google Scholar
• Izaguirre, G. (2019). The proteolytic regulation of virus cell entry by furin and other proprotein convertases. Viruses, 11(9), 837. https://doi.org/10.3390/v11090837
   PubMed Web of Science ®Google Scholar
• Jolliffe, I. T., & Cadima, J. (2016). Principal component analysis: a review and recent developments. Philosophical Transactions. Series A, Mathematical, Physical, and Engineering Sciences, 374(2065), 20150202. https://doi.org/10.1098/rsta.2015.0202
   PubMed Web of Science ®Google Scholar
• Jorgensen, W. L., Chandrasekhar, J., Madura, J. D., Impey, R. W., & Klein, M. L. (1983). Comparison of simple potential functions for simulating liquid water. The Journal of Chemical Physics, 79(2), 926–935. https://doi.org/10.1063/1.445869
   Web of Science ®Google Scholar
• Kadaoluwa Pathirannahalage, S. P., Meftahi, N., Elbourne, A., Weiss, A. C. G., McConville, C. F., Padua, A., Winkler, D. A., Costa Gomes, M., Greaves, T. L., Le, T. C., Besford, Q. A., & Christofferson, A. J. (2021). Systematic comparison of the structural and dynamic properties of commonly used water models for molecular dynamics simulations. Journal of Chemical Information and Modeling, 61(9), 4521–4536. https://doi.org/10.1021/acs.jcim.1c00794
   PubMed Web of Science ®Google Scholar
• Kaminski, G. A., Friesner, R. A., Tirado-Rives, J., & Jorgensen, W. L. (2001). Evaluation and reparametrization of the OPLS-AA force field for proteins via comparison with accurate quantum chemical calculations on peptides. The Journal of Physical Chemistry B, 105(28), 6474–6487. https://doi.org/10.1021/jp003919d
   Web of Science ®Google Scholar
• Laskowski, R. A., MacArthur, M. W., Moss, D. S., & Thornton, J. M. (1993). Procheck: A program to check the stereochemical quality of protein structures. Journal of Applied Crystallography, 26(2), 283–291. https://doi.org/10.1107/S0021889892009944
   Web of Science ®Google Scholar
• Lei, X., Basu, D., Li, Z., Zhang, M., Rudic, R. D., Jiang, X.-C., & Jin, W. (2014). Hepatic overexpression of the prodomain of furin lessens progression of atherosclerosis and reduces vascular remodeling in response to injury. Atherosclerosis, 236(1), 121–130. https://doi.org/10.1016/j.atherosclerosis.2014.06.015
   PubMed Web of Science ®Google Scholar
• Lobanov, M. Y., Bogatyreva, N., & Galzitskaya, O. (2008). Radius of gyration as an indicator of protein structure compactness. Molecular Biology, 42(4), 623–628. https://doi.org/10.1134/S0026893308040195
   Web of Science ®Google Scholar
• Lopes, P. E., Guvench, O., & MacKerell, A. D. (2015). Current status of protein force fields for molecular dynamics simulations. In Molecular modeling of proteins (pp. 47–71). Springer.
   Google Scholar
• MacKerell, A. D., Bashford, D., Bellott, M., Dunbrack, R. L., Evanseck, J. D., Field, M. J., Fischer, S., Gao, J., Guo, H., Ha, S., Joseph-McCarthy, D., Kuchnir, L., Kuczera, K., Lau, F. T., Mattos, C., Michnick, S., Ngo, T., Nguyen, D. T., Prodhom, B., … Karplus, M. (1998). All-atom empirical potential for molecular modeling and dynamics studies of proteins. The Journal of Physical Chemistry. B, 102(18), 3586–3616. https://doi.org/10.1021/jp973084f
   PubMed Web of Science ®Google Scholar
• Mackerell, A. D., Jr Feig, M., & Brooks, III. C. L. (2004). Extending the treatment of backbone energetics in protein force fields: Limitations of gas-phase quantum mechanics in reproducing protein conformational distributions in molecular dynamics simulations. Journal of Computational Chemistry, 25(11), 1400–1415. https://doi.org/10.1002/jcc.20065
   PubMed Web of Science ®Google Scholar
• Madeira, F., Park, Y. M., Lee, J., Buso, N., Gur, T., Madhusoodanan, N., Basutkar, P., Tivey, A. R. N., Potter, S. C., Finn, R. D., & Lopez, R. (2019). The EMBL-EBI search and sequence analysis tools APIs in 2019. Nucleic Acids Research, 47(W1), W636–W641. https://doi.org/10.1093/nar/gkz268
   PubMed Web of Science ®Google Scholar
• Melo, F., & Sali, A. (2007). Fold assessment for comparative protein structure modeling. Protein Science: A Publication of the Protein Society, 16(11), 2412–2426. https://doi.org/10.1110/ps.072895107
   PubMed Web of Science ®Google Scholar
• Messaoudi, A., Belguith, H., & Ben Hamida, J. (2013). Homology modeling and virtual screening approaches to identify potent inhibitors of veb-1 β-lactamase. Theoretical Biology and Medical Modelling, 10(1), 1–10. https://doi.org/10.1186/1742-4682-10-22
   PubMedGoogle Scholar
• Michaud-Agrawal, N., Denning, E. J., Woolf, T. B., & Beckstein, O. (2011). Mdanalysis: A toolkit for the analysis of molecular dynamics simulations. Journal of Computational Chemistry, 32(10), 2319–2327. https://doi.org/10.1002/jcc.21787
   PubMed Web of Science ®Google Scholar
• Mu Y, Kosov DS, Stock G. (2003). Conformational dynamics of trialanine in water. 2. Comparison of amber, charmm, gromos, and OPLs force fields to NMR and infrared experiments. The Journal of Physical Chemistry B, 107(21), 5064–5073. https://doi.org/10.1021/jp022445a
   Web of Science ®Google Scholar
• Needleman, S. B., & Wunsch, C. D. (1970). A general method applicable to the search for similarities in the amino acid sequence of two proteins. Journal of Molecular Biology, 48(3), 443–453. https://doi.org/10.1016/0022-2836(70)90057-4
   PubMed Web of Science ®Google Scholar
• Nowroozi, A., & Shahlaei, M. (2017). A coupling of homology modeling with multiple molecular dynamics simulation for identifying representative conformation of gpcr structures: A case study on human bombesin receptor subtype-3. Journal of Biomolecular Structure & Dynamics, 35(2), 250–272. https://doi.org/10.1080/07391102.2016.1140593
   PubMed Web of Science ®Google Scholar
• Páll, S., & Hess, B. (2013). A flexible algorithm for calculating pair interactions on simd architectures. Computer Physics Communications, 184(12), 2641–2650. https://doi.org/10.1016/j.cpc.2013.06.003
   Web of Science ®Google Scholar
• Pandas Development Team T. (2020). pandas-dev/pandas: Pandas. https://doi.org/10.5281/zenodo.3509134
   Google Scholar
• Parrinello, M., & Rahman, A. (1980). Crystal structure and pair potentials: A molecular-dynamics study. Physical Review Letters, 45(14), 1196–1199. https://doi.org/10.1103/PhysRevLett.45.1196
   Web of Science ®Google Scholar
• Parrinello, M., & Rahman, A. (1981). Polymorphic transitions in single crystals: A new molecular dynamics method. Journal of Applied Physics, 52(12), 7182–7190. https://doi.org/10.1063/1.328693
   Web of Science ®Google Scholar
• Satyanarayana, S. D. V., Krishna, M. S. R., Pavan Kumar, P., & Jeereddy, S. (2018). In silico structural homology modeling of NIF a protein of rhizobial strains in selective legume plants. Journal, Genetic Engineering & Biotechnology, 16(2), 731–737. https://doi.org/10.1016/j.jgeb.2018.06.006
   PubMedGoogle Scholar
• Scamuffa, N., Seidah, N., Basak, A., Cavin, F., Khatib, A. M. (2006). Inactivation of furin by its naturally occurring inhibitor pro-furin abolish breast cancer cells malignant phenotypes and tumorigenecity in mice. Cancer Research, 66(8_Supplement), 1235–1236.
   Web of Science ®Google Scholar
• Schrödinger, L., & DeLano. W. (n.d.). Pymol. http://www.pymol.org/pymol.
   Google Scholar
• Seidah, N. G., Sadr, M. S., Chrétien, M., & Mbikay, M. (2013). The multifaceted proprotein convertases: Their unique, redundant, complementary, and opposite functions. The Journal of Biological Chemistry, 288(30), 21473–21481. https://doi.org/10.1074/jbc.R113.481549
   PubMed Web of Science ®Google Scholar
• Shen, M.-Y., & Sali, A. (2006). Statistical potential for assessment and prediction of protein structures. Protein Science: A Publication of the Protein Society, 15(11), 2507–2524. https://doi.org/10.1110/ps.062416606
   PubMed Web of Science ®Google Scholar
• Sievers, F., Wilm, A., Dineen, D., Gibson, T. J., Karplus, K., Li, W., Lopez, R., McWilliam, H., Remmert, M., Söding, J., Thompson, J. D., & Higgins, D. G. (2011). Fast, scalable generation of high-quality protein multiple sequence alignments using clustal omega. Molecular Systems Biology, 7(1), 539. https://doi.org/10.1038/msb.2011.75
   PubMed Web of Science ®Google Scholar
• Sinha, S., Harioudh, M. K., Dewangan, R. P., Ng, W. J., Ghosh, J. K., & Bhattacharjya, S. (2018). Cell-selective pore forming antimicrobial peptides of the prodomain of human furin: A conserved aromatic/cationic sequence mapping, membrane disruption, and atomic-resolution structure and dynamics. ACS Omega. 3(11), 14650–14664. https://doi.org/10.1021/acsomega.8b01876
   PubMed Web of Science ®Google Scholar
• Tangrea, M. A., Bryan, P. N., Sari, N., & Orban, J. (2002). Solution structure of the pro-hormone convertase 1 pro-domain from mus musculus. Journal of Molecular Biology, 320(4), 801–812. https://doi.org/10.1016/s0022-2836(02)00543-0
   PubMed Web of Science ®Google Scholar
• The GIMP Development Team. (n.d.). Gimp. https://www.gimp.org.
   Google Scholar
• Thomas, G. (2002). Furin at the cutting edge: From protein traffic to embryogenesis and disease. Nature Reviews. Molecular Cell Biology, 3(10), 753–766. https://doi.org/10.1038/nrm934
   PubMed Web of Science ®Google Scholar
• Uniprot: The universal protein knowledgebase. (2021). Nucleic Acids Research, 49(D1), D480–D489.
   PubMed Web of Science ®Google Scholar
• Van Der Spoel, D., Lindahl, E., Hess, B., Groenhof, G., Mark, A. E., & Berendsen, H. J. C. (2005). Gromacs: Fast, flexible, and free. Journal of Computational Chemistry, 26(16), 1701–1718. https://doi.org/10.1002/jcc.20291
   PubMed Web of Science ®Google Scholar
• Waskom, M. L. (2021). Seaborn: Statistical data visualization. Journal of Open Source Software, 6(60), 3021. https://doi.org/10.21105/joss.03021
   Google Scholar
• Webb, B., & Sali, A. (2016). Comparative protein structure modeling using modeller. Current Protocols in Bioinformatics, 54(1), 5–6. https://doi.org/10.1002/cpbi.3
   PubMedGoogle Scholar
• Westerlund, A. M., & Delemotte, L. (2019). Inflecs: Clustering free energy landscapes with Gaussian mixtures. Journal of Chemical Theory and Computation, 15(12), 6752–6759. https://doi.org/10.1021/acs.jctc.9b00454
   PubMed Web of Science ®Google Scholar
• Williams, C. J., Headd, J. J., Moriarty, N. W., Prisant, M. G., Videau, L. L., Deis, L. N., Verma, V., Keedy, D. A., Hintze, B. J., Chen, V. B., Jain, S., Lewis, S. M., Arendall, W. B., Snoeyink, J., Adams, P. D., Lovell, S. C., Richardson, J. S., & Richardson, D. C. (2018). Molprobity: More and better reference data for improved all-atom structure validation. Protein Science: A Publication of the Protein Society, 27(1), 293–315. https://doi.org/10.1002/pro.3330
   PubMed Web of Science ®Google Scholar
• Williamson, D. M., Elferich, J., Ramakrishnan, P., Thomas, G., & Shinde, U. (2013). The mechanism by which a propeptide-encoded ph sensor regulates spatiotemporal activation of furin. The Journal of Biological Chemistry, 288(26), 19154–19165. https://doi.org/10.1074/jbc.M112.442681
   PubMed Web of Science ®Google Scholar
• Winter, A. (2017). QtGrace. Version 0.2.6. https://sourceforge.net/projects/qtgrace/
   Google Scholar
• 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. (2020). A new coronavirus associated with human respiratory disease in china. Nature, 579(7798), 265–269. https://doi.org/10.1038/s41586-020-2008-3
   PubMed Web of Science ®Google Scholar