{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T05:01:56Z","timestamp":1787029316893,"version":"build-2736575974"},"reference-count":40,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2021,9,15]],"date-time":"2021-09-15T00:00:00Z","timestamp":1631664000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001411","name":"Indian Council of Medical Research","doi-asserted-by":"publisher","award":["3\/1\/3\/PDF(8)\/2013-HRD"],"award-info":[{"award-number":["3\/1\/3\/PDF(8)\/2013-HRD"]}],"id":[{"id":"10.13039\/501100001411","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Visceral leishmaniasis (VL), mainly caused by the Leishmania donovani parasitic infection, constitutes a potentially fatal disease, for which treatment is primarily dependent on chemotherapy. The emergence of a resistant parasite towards current antileishmanial agents and increasing reports of relapses are the major concerns. Detailed research on the molecular interaction at the host-parasite interface may provide the identification of the parasite and the host-related factors operating during disease development. Genomic and proteomic studies highlighted several essential secretory and cytosolic proteins that play vital roles during Leishmania pathogenesis. The aim of this study was to identify membrane proteins from the Leishmania donovani parasite and the host macrophage that interact with each other using 2-DE\/MALDI-TOF\/MS. We identified membrane proteins including activated protein C kinase, peroxidoxin, small myristoylated protein 1 (SMP-1), and cytochrome C oxidase from the parasite, while identifying filamin A interacting protein 1(FILIP1) and \u03b2-actin from macrophages. We further investigated parasite replication and persistence within macrophages following the macrophage-amastigote model in the presence or absence of withaferin (WA), an inhibitor of activated C kinase. WA significantly reduced Leishmania donovani replication within host macrophages. This study sheds light on the important interacting proteins for parasite proliferation and virulence, and the establishment of infection within host cells, which can be targeted further to develop a strategy for chemotherapeutic intervention.<\/jats:p>","DOI":"10.3390\/pathogens10091194","type":"journal-article","created":{"date-parts":[[2021,9,14]],"date-time":"2021-09-14T23:34:11Z","timestamp":1631662451000},"page":"1194","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Proteomic Analysis of Leishmania donovani Membrane Components Reveals the Role of Activated Protein C Kinase in Host-Parasite Interaction"],"prefix":"10.3390","volume":"10","author":[{"given":"Sandeep","family":"Verma","sequence":"first","affiliation":[{"name":"ICMR-National Institute of Pathology, Safdarjung Hospital Campus, New Delhi 110029, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deepak Kumar","family":"Deep","sequence":"additional","affiliation":[{"name":"ICMR-National Institute of Pathology, Safdarjung Hospital Campus, New Delhi 110029, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Poonam","family":"Gautam","sequence":"additional","affiliation":[{"name":"ICMR-National Institute of Pathology, Safdarjung Hospital Campus, New Delhi 110029, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8094-4703","authenticated-orcid":false,"given":"Ruchi","family":"Singh","sequence":"additional","affiliation":[{"name":"ICMR-National Institute of Pathology, Safdarjung Hospital Campus, New Delhi 110029, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Poonam","family":"Salotra","sequence":"additional","affiliation":[{"name":"ICMR-National Institute of Pathology, Safdarjung Hospital Campus, New Delhi 110029, India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,9,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"LaMotte, S., Sp\u00e4th, G.F., Rachidi, N., and Prina, E. (2017). The enemy within: Targeting host\u2013parasite interaction for antileishmanial drug discovery. PLoS Negl. Trop. Dis., 11.","DOI":"10.1371\/journal.pntd.0005480"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"371","DOI":"10.1080\/14789450.2018.1468754","article-title":"Understanding Leishmania parasites through proteomics and implications for the clinic","volume":"15","author":"Sundar","year":"2018","journal-title":"Expert Rev. Proteom."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1016\/j.pt.2014.12.012","article-title":"The past, present, and future of Leishmania genomics and transcriptomics","volume":"31","author":"Cantacessi","year":"2015","journal-title":"Trends Parasitol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1590\/S1413-86702006000100001","article-title":"Comparative two-dimensional gel electrophoresis maps for promastigotes of Leishmania amazonensis and Leishmania major","volume":"10","author":"Brobey","year":"2006","journal-title":"Braz. J. Infect. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1016\/j.jprot.2013.02.010","article-title":"Differential quantitative proteomic profiling of Leishmania infantum and Leishmania mexicana density gradient separated membranous fractions","volume":"82","author":"Lynn","year":"2013","journal-title":"J. Proteom."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"e1800136","DOI":"10.1002\/prca.201800136","article-title":"Proteomics and Leishmaniasis: Potential Clinical Applications","volume":"13","author":"Mule","year":"2019","journal-title":"Proteom. Clin. Appl."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"489","DOI":"10.1586\/14760584.2014.894467","article-title":"Identifying vaccine targets for anti-leishmanial vaccine development","volume":"13","author":"Sundar","year":"2014","journal-title":"Expert Rev. Vaccines"},{"key":"ref_8","first-page":"83","article-title":"The early interaction of Leishmania with macrophages and dendritic cells and its influence on the host immune response","volume":"12","author":"Liu","year":"2012","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"4902","DOI":"10.4049\/jimmunol.172.8.4902","article-title":"A Leucine-Rich Repeat Motif of Leishmania Parasite Surface Antigen 2 Binds to Macrophages through the Complement Receptor 3","volume":"172","author":"Kedzierski","year":"2004","journal-title":"J. Immunol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2993","DOI":"10.1242\/jcs.112.18.2993","article-title":"Leishmania species: Models of intracellular parasitism","volume":"112","author":"Alexander","year":"1999","journal-title":"J. Cell Sci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1111\/j.0105-2896.2004.00190.x","article-title":"Does the Leishmania major paradigm of pathogenesis and protection hold for New World cutaneous leishmaniases or the visceral disease?","volume":"201","author":"Alexander","year":"2004","journal-title":"Immunol. Rev."},{"key":"ref_12","first-page":"511","article-title":"The Leishmania Parasitophorous Vacuole Membrane at the Parasite-Host Interface","volume":"92","author":"Young","year":"2019","journal-title":"Yale J. Biol. Med"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"842","DOI":"10.1242\/jcs.056465","article-title":"An exosome-based secretion pathway is responsible for protein export from Leishmania and communication with macrophages","volume":"123","author":"Silverman","year":"2010","journal-title":"J. Cell Sci."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1007\/s004300100076","article-title":"Leishmania infection and virulence","volume":"190","author":"Matlashewski","year":"2001","journal-title":"Med. Microbiol. Immunol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"103464","DOI":"10.1016\/j.jprot.2019.103464","article-title":"Quantitative secretome analysis unravels new secreted proteins in Amphotericin B resistant Leishmania donovani","volume":"207","author":"Garg","year":"2019","journal-title":"J. Proteom."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"111","DOI":"10.2217\/fmb.14.103","article-title":"Leishmania and the macrophage: A multifaceted interaction","volume":"10","author":"Podinovskaia","year":"2015","journal-title":"Futur. Microbiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1002\/biof.5520170110","article-title":"Is there a role of glutathione peroxidases in signaling and differentiation?","volume":"17","year":"2003","journal-title":"BioFactors"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41598-018-30108-2","article-title":"Analyses of the three 1-Cys Peroxiredoxins from Aspergillus fumigatus reveal that cytosolic Prx1 is central to H2O2 metabolism and virulence","volume":"8","author":"Rocha","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"839","DOI":"10.3389\/fimmu.2017.00839","article-title":"l-Arginine uptake by cationic amino acid transporter promotes intra-macrophage survival of Leishmania donovani by enhancing arginase-mediated polyamine synthesis","volume":"8","author":"Mandal","year":"2017","journal-title":"Front. Immunol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1186\/1559-0275-10-9","article-title":"Characterizing the normal proteome of human ciliary body","volume":"10","author":"Goel","year":"2013","journal-title":"Clin. Proteom."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1016\/j.parint.2015.01.004","article-title":"Proteomic analyses of membrane enriched proteins of Leishmania donovani Indian clinical isolate by mass spectrometry","volume":"64","author":"Kumar","year":"2015","journal-title":"Parasitol. Int."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"3289","DOI":"10.1016\/j.bbagen.2012.11.020","article-title":"Glutathione peroxidases","volume":"1830","author":"Maiorino","year":"2013","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"350","DOI":"10.1002\/pmic.200700697","article-title":"Phosphoproteomic analysis of Leishmania donovani pro- and amastigote stages","volume":"8","author":"Morales","year":"2008","journal-title":"Proteomics"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"608","DOI":"10.1089\/ars.2011.4404","article-title":"Peroxiredoxins in Parasites","volume":"17","author":"Gretes","year":"2012","journal-title":"Antioxid. Redox Signal."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"4775","DOI":"10.1091\/mbc.e04-06-0457","article-title":"SMP-1, a Member of a New Family of Small Myristoylated Proteins in Kinetoplastid Parasites, Is Targeted to the Flagellum Membrane in Leishmania","volume":"15","author":"Tull","year":"2004","journal-title":"Mol. Biol. Cell"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Taylor, C.M., Fischer, K., Abubucker, S., Wang, Z., Martin, J., Jiang, D., Magliano, M., Rosso, M.-N., Li, B.-W., and Fischer, P.U. (2011). Targeting Protein-Protein Interactions for Parasite Control. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0018381"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"492","DOI":"10.3389\/fimmu.2019.00492","article-title":"Leishmania-Host Interactions\u2014An Epigenetic Paradigm","volume":"10","author":"Afrin","year":"2019","journal-title":"Front. Immunol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1155\/2012\/819512","article-title":"Host Cell Signalling and Leishmania: Mechanisms of Evasion","volume":"2012","author":"Shio","year":"2012","journal-title":"J. Trop. Med."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1689","DOI":"10.1084\/jem.20031162","article-title":"Leishmania major LACK Antigen Is Required for Efficient Vertebrate Parasitization","volume":"198","author":"Kelly","year":"2003","journal-title":"J. Exp. Med."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1111\/mmi.12924","article-title":"LACK, a RACK1 ortholog, facilitates cytochrome c oxidase subunit expression to promote Leishmania major fitness","volume":"96","author":"Cardenas","year":"2015","journal-title":"Mol. Microbiol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1287","DOI":"10.1007\/s00436-009-1554-2","article-title":"Evaluation of anti-leishmanial activity of selected Indian plants known to have antimicrobial properties","volume":"105","author":"Sharma","year":"2009","journal-title":"Parasitol. Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1029","DOI":"10.3109\/14756366.2015.1088841","article-title":"Exploring the inhibitory activity of Withaferin-A against Pteridine reductase-1 of L. donovani","volume":"31","author":"Chandrasekaran","year":"2015","journal-title":"J. Enzym. Inhib. Med. Chem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2011","DOI":"10.1016\/j.bbamem.2014.04.017","article-title":"Integrity of the Actin Cytoskeleton of Host Macrophages is Essential for Leishmania donovani Infection","volume":"1838","author":"Roy","year":"2014","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1186\/2045-3701-3-7","article-title":"Complex roles of filamin-A mediated cytoskeleton network in cancer progression","volume":"3","author":"Yue","year":"2013","journal-title":"Cell Biosci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"825","DOI":"10.1007\/s00436-012-3212-3","article-title":"Validation of a simple resazurin-based promastigote assay for the routine monitoring of miltefosine susceptibility in clinical isolates of Leishmania donovani","volume":"112","author":"Kulshrestha","year":"2013","journal-title":"Parasitol. Res."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"835","DOI":"10.1128\/AAC.01233-08","article-title":"In Vitro Susceptibility of Field Isolates of Leishmania donovani to Miltefosine and Amphotericin B: Correlation with Sodium Antimony Gluconate Susceptibility and Implications for Treatment in Areas of Endemicity","volume":"53","author":"Kumar","year":"2008","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"5197","DOI":"10.1128\/AAC.00866-09","article-title":"In Vitro Sensitivity Testing of Leishmania Clinical Field Isolates: Preconditioning of Promastigotes Enhances Infectivity for Macrophage Host Cells","volume":"53","author":"Vermeersch","year":"2009","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1016\/S0925-4439(99)00034-4","article-title":"Receptor\/ligand interactions between Cryptosporidium parvum and the surface of the host cell","volume":"1454","author":"Nesterenko","year":"1999","journal-title":"Biochim. Biophys. Acta (BBA) Mol. Basis Dis."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"23603","DOI":"10.1074\/jbc.M115.645341","article-title":"Adipose Tissue Lipolysis Promotes Exercise-induced Cardiac Hypertrophy Involving the Lipokine C16:1n7-Palmitoleate","volume":"290","author":"Kreissl","year":"2015","journal-title":"J. Biol. Chem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1568","DOI":"10.1016\/j.fct.2010.03.026","article-title":"Supercritical fluid extraction of oregano (Origanum vulgare) essentials oils: Anti-inflammatory properties based on cytokine response on THP-1 macrophages","volume":"48","author":"Reglero","year":"2010","journal-title":"Food Chem. Toxicol."}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/10\/9\/1194\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:02:55Z","timestamp":1760166175000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/10\/9\/1194"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,9,15]]},"references-count":40,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2021,9]]}},"alternative-id":["pathogens10091194"],"URL":"https:\/\/doi.org\/10.3390\/pathogens10091194","relation":{},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,9,15]]}}}