{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,12]],"date-time":"2025-11-12T14:15:17Z","timestamp":1762956917945,"version":"build-2065373602"},"reference-count":48,"publisher":"MDPI AG","issue":"14","license":[{"start":{"date-parts":[[2022,7,20]],"date-time":"2022-07-20T00:00:00Z","timestamp":1658275200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001807","name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de S\u00e3o Paulo-FAPESP","doi-asserted-by":"publisher","award":["2017\/08834-9"],"award-info":[{"award-number":["2017\/08834-9"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Federal Institute of Education, Science and Technology of Mato Grosso (IFMT)","award":["2017\/08834-9"],"award-info":[{"award-number":["2017\/08834-9"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Herein, we elucidate the biophysical aspects of the interaction of an important protein, Interleukin-6 (IL6), which is involved in cytokine storm syndrome, with a natural product with anti-inflammatory activity, piperine. Despite the role of piperine in the inhibition of the transcriptional protein NF-\u03baB pathway responsible for activation of IL6 gene expression, there are no studies to the best of our knowledge regarding the characterisation of the molecular interaction of the IL6-piperine complex. In this context, the characterisation was performed with spectroscopic experiments aided by molecular modelling. Fluorescence spectroscopy alongside van\u2019t Hoff analyses showed that the complexation event is a spontaneous process driven by non-specific interactions. Circular dichroism aided by molecular dynamics revealed that piperine caused local \u03b1-helix reduction. Molecular docking and molecular dynamics disclosed the microenvironment of interaction as non-polar amino acid residues. Although piperine has three available hydrogen bond acceptors, only one hydrogen-bond was formed during our simulation experiments, reinforcing the major role of non-specific interactions that we observed experimentally. Root mean square deviation (RMSD) and hydrodynamic radii revealed that the IL6-piperine complex was stable during 800 ns of simulation. Taken together, these results can support ongoing IL6 drug discovery efforts.<\/jats:p>","DOI":"10.3390\/ijms23147994","type":"journal-article","created":{"date-parts":[[2022,7,20]],"date-time":"2022-07-20T11:22:24Z","timestamp":1658316144000},"page":"7994","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["A Computational\u2013Experimental Investigation of the Molecular Mechanism of Interleukin-6-Piperine Interaction"],"prefix":"10.3390","volume":"23","author":[{"given":"Ana Paula Ribeiro","family":"Povinelli","sequence":"first","affiliation":[{"name":"Federal Institute of Education, Science and Technology of Mato Grosso, Campo Novo do Parecis 78360-000, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gabriel","family":"Zazeri","sequence":"additional","affiliation":[{"name":"Federal Institute of Education, Science and Technology of Mato Grosso, Campo Novo do Parecis 78360-000, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3897-5626","authenticated-orcid":false,"given":"Alan M.","family":"Jones","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Institute of Clinical Sciences, College of Medical and Dental Sciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0755-8643","authenticated-orcid":false,"given":"Marin\u00f4nio Lopes","family":"Corn\u00e9lio","sequence":"additional","affiliation":[{"name":"Departamento de F\u00edsica, Instituto de Bioci\u00eancias, Letras e Ci\u00eancias Exatas (IBILCE), UNESP, Rua Cristov\u00e3o Colombo 2265, S\u00e3o Jos\u00e9 do Rio Preto 15054-000, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,7,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"S2","DOI":"10.1186\/ar1916","article-title":"Interleukin-6: Discovery of a pleiotropic cytokine","volume":"8","author":"Kishimoto","year":"2006","journal-title":"Arthritis Res. Ther."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/j.pharmthera.2013.09.004","article-title":"Targeting interleukin-6 in inflammatory autoimmune diseases and cancers","volume":"141","author":"Yao","year":"2014","journal-title":"Pharmacol. Ther."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1007\/978-1-4613-1241-3_8","article-title":"Interleukin-6: A comprehensive review","volume":"80","author":"Lotz","year":"1995","journal-title":"Cytokines Interleukins Recept."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"655","DOI":"10.1016\/S2213-2600(21)00139-9","article-title":"Interleukin-6 receptor blockade in patients with COVID-19: Placing clinical trials into context","volume":"9","author":"Angriman","year":"2021","journal-title":"Lancet Respir. Med."},{"key":"ref_5","first-page":"2327","article-title":"Activation of interleukin-6 gene expression through the NF-kappa B transcription factor","volume":"10","author":"Libermann","year":"1990","journal-title":"Mol. Cell. Biol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"e13","DOI":"10.1016\/S2213-2600(20)30600-7","article-title":"Assessing the importance of interleukin-6 in COVID-19","volume":"9","author":"Chen","year":"2021","journal-title":"Lancet Respir. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"110698","DOI":"10.1016\/j.biopha.2020.110698","article-title":"The role of Interleukin 6 inhibitors in therapy of severe COVID-19","volume":"131","author":"Nasonov","year":"2020","journal-title":"Biomed. Pharmacother."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1717","DOI":"10.4014\/jmb.1604.04051","article-title":"Enhancing the thermal resistance of a novel acidobacteria-derived phytase by engineering of disulfide bridges","volume":"26","author":"Tan","year":"2016","journal-title":"J. Microbiol. Biotechnol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1016\/j.enzmictec.2013.08.001","article-title":"Significant improvement of thermal stability of glucose 1-dehydrogenase by introducing disulfide bonds at the tetramer interface","volume":"53","author":"Ding","year":"2013","journal-title":"Enzyme Microb. Technol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"989","DOI":"10.1093\/emboj\/16.5.989","article-title":"1.9 \u00c5 crystal structure of interleukin 6: Implications for a novel mode of receptor dimerization and signaling","volume":"16","author":"Somers","year":"1997","journal-title":"EMBO J."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"468","DOI":"10.1006\/jmbi.1997.0933","article-title":"Solution structure of recombinant human interleukin-6","volume":"268","author":"Xu","year":"1997","journal-title":"J. Mol. Biol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"155684","DOI":"10.1016\/j.cyto.2021.155684","article-title":"Unravelling the broader complexity of IL-6 involvement in health and disease","volume":"148","author":"Jenkins","year":"2021","journal-title":"Cytokine"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"641","DOI":"10.2174\/1573409915666190902142524","article-title":"In silico insights on IL-6: A potential target for multicentric castleman disease","volume":"16","author":"Aher","year":"2020","journal-title":"Curr. Comput. Aided. Drug Des."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1121","DOI":"10.1002\/ptr.4972","article-title":"Piper nigrum and piperine: An update","volume":"27","author":"Meghwal","year":"2013","journal-title":"Phyther. Res."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Zazeri, G., Povinelli, A.P.R., Le Duff, C.S., Tang, B., Cornelio, M.L., and Jones, A.M. (2020). Synthesis and Spectroscopic Analysis of Piperine-and Piperlongumine-Inspired Natural Product Scaffolds and Their Molecular Docking with IL-1\u03b2 and NF-\u03baB Proteins. Molecules, 25.","DOI":"10.3390\/molecules25122841"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1016\/j.intimp.2013.06.025","article-title":"Piperine inhibits IL-\u03b2 induced expression of inflammatory mediators in human osteoarthritis chondrocyte","volume":"17","author":"Ying","year":"2013","journal-title":"Int. Immunopharmacol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/j.cellimm.2013.09.001","article-title":"Piperine inhibits LPS induced expression of inflammatory mediators in RAW 264.7 cells","volume":"285","author":"Ying","year":"2013","journal-title":"Cell. Immunol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"R49","DOI":"10.1186\/ar2662","article-title":"Anti-inflammatory and antiarthritic effects of piperine in human interleukin 1\u03b2-stimulated fibroblast-like synoviocytes and in rat arthritis models","volume":"11","author":"Bang","year":"2009","journal-title":"Arthritis Res. Ther."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"10179","DOI":"10.1021\/jf051944g","article-title":"Binding of the pepper alkaloid piperine to bovine \u03b2-lactoglobulin: Circular dichroism spectroscopy and molecular modeling study","volume":"53","author":"Zsila","year":"2005","journal-title":"J. Agric. Food Chem."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Zazeri, G., Povinelli, A.P.R., Lima, M.D.F., and Corn\u00e9lio, M.L. (2020). The Cytokine IL-1\u03b2 and Piperine Complex Surveyed by Experimental and Computational Molecular Biophysics. Biomolecules, 10.","DOI":"10.3390\/biom10091337"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1186\/s43088-022-00196-1","article-title":"Molecular and pharmacological aspects of piperine as a potential molecule for disease prevention and management: Evidence from clinical trials","volume":"11","author":"Tripathi","year":"2022","journal-title":"Beni-Suef Univ. J. Basic Appl. Sci."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Zazeri, G., Povinelli, A.P.R., Lima, M.D.F., and Corn\u00e9lio, M.L. (2020). Detailed Characterization of the Cooperative Binding of Piperine with Heat Shock Protein 70 by Molecular Biophysical Approaches. Biomedicines, 8.","DOI":"10.3390\/biomedicines8120629"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"15667","DOI":"10.1038\/s41598-019-52187-5","article-title":"Details of the cooperative binding of piperlongumine with rat serum albumin obtained by spectroscopic and computational analyses","volume":"9","author":"Povinelli","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Povinelli, A.P.R., Zazeri, G., Jones, A.M., and Corn\u00e9lio, M.L. (2021). Unravelling the Interaction of Piperlongumine with the Nucleotide-Binding Domain of HSP70: A Spectroscopic and In Silico Study. Pharmaceuticals, 14.","DOI":"10.3390\/ph14121298"},{"key":"ref_25","unstructured":"Povinelli, A.P.R., Zazeri, G., and Corn\u00e9lio, M.L. (2019). Molecular Mechanism of Flavonoids Using Fluorescence Spectroscopy and Computational Tools. Flavonoids-A Coloring Model For Cheering Up Life, IntechOpen."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Lakowicz, J.R. (2004). Principles of Fluorescence Spectroscopy, (1999), Kluwer Academic\/Plenum Publishers.","DOI":"10.1007\/978-1-4757-3061-6"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"6726","DOI":"10.1021\/jf070905x","article-title":"Interaction of different polyphenols with bovine serum albumin (BSA) and human salivary \u03b1-amylase (HSA) by fluorescence quenching","volume":"55","author":"Soares","year":"2007","journal-title":"J. Agric. Food Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"3096","DOI":"10.1021\/bi00514a017","article-title":"Thermodynamics of protein association reactions: Forces contributing to stability","volume":"20","author":"Ross","year":"1981","journal-title":"Biochemistry"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Zazeri, G., Povinelli, A.P.R., Lima, M.D.F., and Corn\u00e9lio, M.L. (2019). Experimental approaches and computational modeling of rat serum albumin and its interaction with piperine. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20122856"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"130995","DOI":"10.1016\/j.molstruc.2021.130995","article-title":"Experimental studies and computational modeling on cytochrome c reduction by quercetin: The role of oxidability and binding affinity","volume":"1244","author":"Zazeri","year":"2021","journal-title":"J. Mol. Struct."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1016\/0014-5793(90)80219-9","article-title":"Structure-function analysis of human interleukin-6. Evidence for the involvement of the carboxy-terminus in function","volume":"262","author":"Wroblowski","year":"1990","journal-title":"FEBS Lett."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"903","DOI":"10.1111\/j.1432-1033.1992.tb17123.x","article-title":"Purification and characterization of a recombinant murine interleukin-6: Isolation of N- and C-terminally truncated forms","volume":"207","author":"Zhang","year":"1992","journal-title":"Eur. J. Biochem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"4120","DOI":"10.1021\/bi00772a015","article-title":"Determination of the secondary structures of proteins by circular dichroism and optical rotatory dispersion","volume":"11","author":"Chen","year":"1972","journal-title":"Biochemistry"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1002\/pro.3280","article-title":"Improvements to the APBS biomolecular solvation software suite","volume":"27","author":"Jurrus","year":"2018","journal-title":"Protein Sci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2785","DOI":"10.1002\/jcc.21256","article-title":"AutoDock4 and AutoDockTools4: Automated docking with selective receptor flexibility","volume":"30","author":"Morris","year":"2009","journal-title":"J. Comput. Chem."},{"key":"ref_36","unstructured":"Morris, G.M., Goodsell, D.S., Pique, M.E., Lindstrom, W.L., Huey, R., Forli, S., Hart, W.E., Halliday, S., Belew, R., and Olson, A.J. User Guide AutoDock version 4.2. Automated Docking of Flexible Ligands to Flexible Receptors."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/0263-7855(96)00018-5","article-title":"VMD: Visual molecular dynamics","volume":"14","author":"Humphrey","year":"1996","journal-title":"J. Mol. Graph."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1093\/protein\/8.2.127","article-title":"LIGPLOT: A program to generate schematic diagrams of protein-ligand interactions","volume":"8","author":"Wallace","year":"1995","journal-title":"Protein Eng. Des. Sel."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1656","DOI":"10.1002\/jcc.20090","article-title":"A biomolecular force field based on the free enthalpy of hydration and solvation: The GROMOS force-field parameter sets 53A5 and 53A6","volume":"25","author":"Oostenbrink","year":"2004","journal-title":"J. Comput. Chem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1701","DOI":"10.1002\/jcc.20291","article-title":"GROMACS: Fast, flexible, and free","volume":"26","author":"Lindahl","year":"2005","journal-title":"J. Comput. Chem."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"4026","DOI":"10.1021\/ct200196m","article-title":"An automated force field topology builder (ATB) and repository: Version 1.0","volume":"7","author":"Malde","year":"2011","journal-title":"J. Chem. Theory Comput."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"24503","DOI":"10.1063\/1.2136877","article-title":"Flexible simple point-charge water model with improved liquid-state properties","volume":"124","author":"Wu","year":"2006","journal-title":"J. Chem. Phys."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"14101","DOI":"10.1063\/1.2408420","article-title":"Canonical sampling through velocity rescaling","volume":"126","author":"Bussi","year":"2007","journal-title":"J. Chem. Phys."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1463","DOI":"10.1002\/(SICI)1096-987X(199709)18:12<1463::AID-JCC4>3.0.CO;2-H","article-title":"LINCS: A linear constraint solver for molecular simulations","volume":"18","author":"Hess","year":"1997","journal-title":"J. Comput. Chem."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"4003","DOI":"10.1063\/1.1389854","article-title":"Optimized particle-mesh Ewald\/multiple-time step integration for molecular dynamics simulations","volume":"115","author":"Batcho","year":"2001","journal-title":"J. Chem. Phys."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"7182","DOI":"10.1063\/1.328693","article-title":"Polymorphic transitions in single crystals: A new molecular dynamics method","volume":"52","author":"Parrinello","year":"1981","journal-title":"J. Appl. Phys."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1016\/0010-4655(95)00053-I","article-title":"The calculation of the potential of mean force using computer simulations","volume":"91","author":"Roux","year":"1995","journal-title":"Comput. Phys. Commun."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1011","DOI":"10.1002\/jcc.540130812","article-title":"The weighted histogram analysis method for free-energy calculations on biomolecules. I. The method","volume":"13","author":"Kumar","year":"1992","journal-title":"J. Comput. Chem."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/14\/7994\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:54:28Z","timestamp":1760140468000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/14\/7994"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,7,20]]},"references-count":48,"journal-issue":{"issue":"14","published-online":{"date-parts":[[2022,7]]}},"alternative-id":["ijms23147994"],"URL":"https:\/\/doi.org\/10.3390\/ijms23147994","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2022,7,20]]}}}