{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,16]],"date-time":"2026-02-16T20:46:58Z","timestamp":1771274818458,"version":"3.50.1"},"reference-count":36,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2024,9,19]],"date-time":"2024-09-19T00:00:00Z","timestamp":1726704000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Nazarbayev University Faculty-Development Competitive Research","award":["20122022FD4123"],"award-info":[{"award-number":["20122022FD4123"]}]},{"name":"Nazarbayev University Faculty-Development Competitive Research","award":["20122022CRP1615"],"award-info":[{"award-number":["20122022CRP1615"]}]},{"name":"Collaborative Research","award":["20122022FD4123"],"award-info":[{"award-number":["20122022FD4123"]}]},{"name":"Collaborative Research","award":["20122022CRP1615"],"award-info":[{"award-number":["20122022CRP1615"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>The collaboration between cellular proteases and host cells is pivotal in mounting an effective innate immune defense. Of particular interest is the synergistic interaction between cathepsin G (CatG) and neutrophil elastase (NE), which are proteases secreted by activated neutrophils, and the human alveolar basal epithelial cell line (A549) and the human lung epithelial-like cell line (H1299), because of the potential implications for viral infection. Our study aimed to investigate the binding capacity of CatG and NE on the surface of A549 and H1299 cells through preincubation with purified CatG and NE; thereby, the proteolytic activity could be detected using activity-based probes. Both CatG and NE were capable of binding to the cell surface and exhibited proteolytic activity, leading to increased cell surface levels of MHC I molecules, which is crucial for displaying the endogenous antigenic repertoire. In addition, CatG cleaved the S2\u2032 site of the SARS-CoV-2 spike protein at two specific sites (815RS816 and 817FI818) as well as NE (813SK814 and 818IE819), which potentially leads to the destruction of the fusion peptide. Additionally, furin required the presence of Ca2+ ions for the distinct cleavage site necessary to generate the fusion peptide. Overall, the findings suggest that CatG and NE can fortify target cells against viral entry, underscoring the potential significance of cell surface proteases in protecting against viral invasion.<\/jats:p>","DOI":"10.3390\/molecules29184449","type":"journal-article","created":{"date-parts":[[2024,9,19]],"date-time":"2024-09-19T08:12:27Z","timestamp":1726733547000},"page":"4449","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["The Proteolytic Activity of Neutrophil-Derived Serine Proteases Bound to the Cell Surface Arming Lung Epithelial Cells for Viral Defense"],"prefix":"10.3390","volume":"29","author":[{"given":"Akmaral","family":"Assylbekova","sequence":"first","affiliation":[{"name":"Department of Biology, School of Sciences and Humanities, Nazarbayev University, Kabanbay Batyr Ave. 53, Astana 010000, Kazakhstan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maiya","family":"Allayarova","sequence":"additional","affiliation":[{"name":"Department of Biology, School of Sciences and Humanities, Nazarbayev University, Kabanbay Batyr Ave. 53, Astana 010000, Kazakhstan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Moldir","family":"Konysbekova","sequence":"additional","affiliation":[{"name":"Department of Biology, School of Sciences and Humanities, Nazarbayev University, Kabanbay Batyr Ave. 53, Astana 010000, Kazakhstan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Amanbek","family":"Bekturgan","sequence":"additional","affiliation":[{"name":"Department of Biology, School of Sciences and Humanities, Nazarbayev University, Kabanbay Batyr Ave. 53, Astana 010000, Kazakhstan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aiya","family":"Makhanova","sequence":"additional","affiliation":[{"name":"Department of Biology, School of Sciences and Humanities, Nazarbayev University, Kabanbay Batyr Ave. 53, Astana 010000, Kazakhstan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5001-525X","authenticated-orcid":false,"given":"Samantha","family":"Brown","sequence":"additional","affiliation":[{"name":"Institute for Archaeological Sciences, Department of Geosciences, University of T\u00fcbingen, 72076 T\u00fcbingen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-7555-3660","authenticated-orcid":false,"given":"Norbert","family":"Grzegorzek","sequence":"additional","affiliation":[{"name":"Mass Spectrometry Facility, Organic Chemistry, Eberhard Karls University T\u00fcbingen, 72076 T\u00fcbingen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hubert","family":"Kalbacher","sequence":"additional","affiliation":[{"name":"Institute of Clinical Anatomy and Cell Analysis, University Hospital T\u00fcbingen, Eberhard Karls University T\u00fcbingen, \u00d6sterbergstra\u00dfe 3, 72074 T\u00fcbingen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3986-2460","authenticated-orcid":false,"given":"Ruslan","family":"Kalendar","sequence":"additional","affiliation":[{"name":"Center for Life Sciences, National Laboratory Astana, Nazarbayev University, Kabanbay Batyr Ave. 53, Astana 010000, Kazakhstan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9596-6558","authenticated-orcid":false,"given":"Timo","family":"Burster","sequence":"additional","affiliation":[{"name":"Department of Biology, School of Sciences and Humanities, Nazarbayev University, Kabanbay Batyr Ave. 53, Astana 010000, Kazakhstan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,9,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"726","DOI":"10.1124\/pr.110.002733","article-title":"Neutrophil elastase, proteinase 3, and cathepsin G as therapeutic targets in human diseases","volume":"62","author":"Korkmaz","year":"2010","journal-title":"Pharmacol. Rev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1943","DOI":"10.1016\/j.molimm.2010.04.020","article-title":"Serine proteases of the human immune system in health and disease","volume":"47","author":"Heutinck","year":"2010","journal-title":"Mol. Immunol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"74602","DOI":"10.18632\/oncotarget.12980","article-title":"Exogenous cathepsin G upregulates cell surface MHC class I molecules on immune and glioblastoma cells","volume":"7","author":"Giese","year":"2016","journal-title":"Oncotarget"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1007\/s00005-020-00585-3","article-title":"Cathepsin G and its Dichotomous Role in Modulating Levels of MHC Class I Molecules","volume":"68","author":"Burster","year":"2020","journal-title":"Arch. Immunol. Ther. Exp."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"741","DOI":"10.1007\/s00262-016-1841-6","article-title":"Neutrophil elastase enhances antigen presentation by upregulating human leukocyte antigen class I expression on tumor cells","volume":"65","author":"Chawla","year":"2016","journal-title":"Cancer Immunol. Immunother."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1158\/2326-6066.CIR-16-0141","article-title":"Serine Proteases Enhance Immunogenic Antigen Presentation on Lung Cancer Cells","volume":"5","author":"Peters","year":"2017","journal-title":"Cancer Immunol. Res."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Wettstein, L., Kirchhoff, F., and Munch, J. (2022). The Transmembrane Protease TMPRSS2 as a Therapeutic Target for COVID-19 Treatment. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23031351"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1038\/s41580-021-00418-x","article-title":"Mechanisms of SARS-CoV-2 entry into cells","volume":"23","author":"Jackson","year":"2022","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"779","DOI":"10.1016\/j.molcel.2020.04.022","article-title":"A Multibasic Cleavage Site in the Spike Protein of SARS-CoV-2 Is Essential for Infection of Human Lung Cells","volume":"78","author":"Hoffmann","year":"2020","journal-title":"Mol. Cell"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1111\/1348-0421.12945","article-title":"Proteolytic activation of SARS-CoV-2 spike protein","volume":"66","author":"Takeda","year":"2022","journal-title":"Microbiol. Immunol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"271","DOI":"10.1016\/j.cell.2020.02.052","article-title":"SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a Clinically Proven Protease Inhibitor","volume":"181","author":"Hoffmann","year":"2020","journal-title":"Cell"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Ou, T., Mou, H., Zhang, L., Ojha, A., Choe, H., and Farzan, M. (2021). Hydroxychloroquine-mediated inhibition of SARS-CoV-2 entry is attenuated by TMPRSS2. PLoS Pathog., 17.","DOI":"10.1371\/journal.ppat.1009212"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"e174133","DOI":"10.1172\/jci.insight.174133","article-title":"Neutrophil proteases are protective against SARS-CoV-2 by degrading the spike protein and dampening virus-mediated inflammation","volume":"9","author":"Leborgne","year":"2024","journal-title":"JCI Insight"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1016\/j.biochi.2007.10.009","article-title":"Neutrophil elastase, proteinase 3 and cathepsin G: Physicochemical properties, activity and physiopathological functions","volume":"90","author":"Korkmaz","year":"2008","journal-title":"Biochimie"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Schroeder, R., Grzywa, R., Wirtz, C.R., Sienczyk, M., and Burster, T. (2020). Application of a novel FAM-conjugated activity-based probe to determine cathepsin G activity intracellularly. Anal. Biochem., 588.","DOI":"10.1016\/j.ab.2019.113488"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2605","DOI":"10.1002\/cbic.201402360","article-title":"Synthesis of novel phosphonic-type activity-based probes for neutrophil serine proteases and their application in spleen lysates of different organisms","volume":"15","author":"Grzywa","year":"2014","journal-title":"Chembiochem"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Burster, T., Gartner, F., Knippschild, U., and Zhanapiya, A. (2021). Activity-Based Probes to Utilize the Proteolytic Activity of Cathepsin G in Biological Samples. Front. Chem., 9.","DOI":"10.3389\/fchem.2021.628295"},{"key":"ref_18","first-page":"C106","article-title":"An integrative view of serpins in health and disease: The contribution of SerpinA3","volume":"320","author":"Bobadilla","year":"2021","journal-title":"Am. J. Physiol. Cell Physiol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"963","DOI":"10.1038\/s41589-022-01059-7","article-title":"Structure and activity of human TMPRSS2 protease implicated in SARS-CoV-2 activation","volume":"18","author":"Fraser","year":"2022","journal-title":"Nat. Chem. Biol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"7181","DOI":"10.1021\/acsomega.1c00363","article-title":"Neutrophil Elastase and Proteinase 3 Cleavage Sites Are Adjacent to the Polybasic Sequence within the Proteolytic Sensitive Activation Loop of the SARS-CoV-2 Spike Protein","volume":"6","author":"Mustafa","year":"2021","journal-title":"ACS Omega"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"e2300644120","DOI":"10.1073\/pnas.2300644120","article-title":"Viral afterlife: SARS-CoV-2 as a reservoir of immunomimetic peptides that reassemble into proinflammatory supramolecular complexes","volume":"121","author":"Zhang","year":"2024","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Mustafa, Z., Kalbacher, H., and Burster, T. (2022). Occurrence of a novel cleavage site for cathepsin G adjacent to the polybasic sequence within the proteolytically sensitive activation loop of the SARS-CoV-2 Omicron variant: The amino acid substitution N679K and P681H of the spike protein. PLoS ONE, 17.","DOI":"10.1371\/journal.pone.0264723"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1149","DOI":"10.1007\/s43440-022-00394-9","article-title":"The roles of cellular protease interactions in viral infections and programmed cell death: A lesson learned from the SARS-CoV-2 outbreak and COVID-19 pandemic","volume":"74","author":"Majchrzak","year":"2022","journal-title":"Pharmacol. Rep."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"5495","DOI":"10.4049\/jimmunol.172.9.5495","article-title":"Cathepsin G, and not the asparagine-specific endoprotease, controls the processing of myelin basic protein in lysosomes from human B lymphocytes","volume":"172","author":"Burster","year":"2004","journal-title":"J. Immunol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"764","DOI":"10.1038\/ki.2015.159","article-title":"Neutrophil serine proteases exert proteolytic activity on endothelial cells","volume":"88","author":"Jerke","year":"2015","journal-title":"Kidney Int."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"108573","DOI":"10.1016\/j.intimp.2022.108573","article-title":"Serine proteases in neutrophil extracellular traps exhibit anti-Respiratory Syncytial Virus activity","volume":"106","author":"Lopes","year":"2022","journal-title":"Int. Immunopharmacol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"35341","DOI":"10.1074\/jbc.M112.385617","article-title":"Clathrin pit-mediated endocytosis of neutrophil elastase and cathepsin G by cancer cells","volume":"287","author":"Gregory","year":"2012","journal-title":"J. Biol. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"14","DOI":"10.26508\/lsa.202000786","article-title":"TMPRSS2 and furin are both essential for proteolytic activation of SARS-CoV-2 in human airway cells","volume":"3","author":"Bestle","year":"2020","journal-title":"Life Sci. Alliance"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Lotke, R., Petersen, M., and Sauter, D. (2024). Restriction of Viral Glycoprotein Maturation by Cellular Protease Inhibitors. Viruses, 16.","DOI":"10.3390\/v16030332"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Janciauskiene, S., Wrenger, S., Immenschuh, S., Olejnicka, B., Greulich, T., Welte, T., and Chorostowska-Wynimko, J. (2018). The Multifaceted Effects of Alpha1-Antitrypsin on Neutrophil Functions. Front. Pharmacol., 9.","DOI":"10.3389\/fphar.2018.00341"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"3931","DOI":"10.1016\/S0021-9258(19)85615-6","article-title":"Kinetics of association of serine proteinases with native and oxidized alpha-1-proteinase inhibitor and alpha-1-antichymotrypsin","volume":"255","author":"Beatty","year":"1980","journal-title":"J. Biol. Chem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"eabj7521","DOI":"10.1126\/scitranslmed.abj7521","article-title":"Identification of driver genes for critical forms of COVID-19 in a deeply phenotyped young patient cohort","volume":"14","author":"Carapito","year":"2022","journal-title":"Sci. Transl. Med."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Eipper, S., Steiner, R., Lesner, A., Sienczyk, M., Palesch, D., Halatsch, M.E., Zaczynska, E., Heim, C., Hartmann, M.D., and Zimecki, M. (2016). Lactoferrin Is an Allosteric Enhancer of the Proteolytic Activity of Cathepsin G. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0151509"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.clnesp.2021.08.019","article-title":"Lactoferrin reduces the risk of respiratory tract infections: A meta-analysis of randomized controlled trials","volume":"45","author":"Ali","year":"2021","journal-title":"Clin. Nutr. ESPEN"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"463","DOI":"10.1007\/s10534-022-00477-3","article-title":"Bovine lactoferrin for the prevention of COVID-19 infection in health care personnel: A double-blinded randomized clinical trial (LF-COVID)","volume":"36","author":"Navarro","year":"2023","journal-title":"Biometals"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"123","DOI":"10.4049\/jimmunol.133.1.123","article-title":"The role of T3 surface molecules in the activation of human T cells: A two-stimulus requirement for IL 2 production reflects events occurring at a pre-translational level","volume":"133","author":"Weiss","year":"1984","journal-title":"J. Immunol."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/29\/18\/4449\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T15:59:34Z","timestamp":1760111974000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/29\/18\/4449"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,9,19]]},"references-count":36,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2024,9]]}},"alternative-id":["molecules29184449"],"URL":"https:\/\/doi.org\/10.3390\/molecules29184449","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,9,19]]}}}