{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T07:34:41Z","timestamp":1790062481695,"version":"4.0.1"},"reference-count":34,"publisher":"Springer Science and Business Media LLC","issue":"2","license":[{"start":{"date-parts":[[2024,2,1]],"date-time":"2024-02-01T00:00:00Z","timestamp":1706745600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2024,2,21]],"date-time":"2024-02-21T00:00:00Z","timestamp":1708473600000},"content-version":"vor","delay-in-days":20,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"name":"Austrian Research Fund,Austria","award":["P31793"],"award-info":[{"award-number":["P31793"]}]},{"name":"Austrian Research Fund,Austria","award":["P31793"],"award-info":[{"award-number":["P31793"]}]},{"name":"Austrian Research Fund,Austria","award":["P31793"],"award-info":[{"award-number":["P31793"]}]},{"DOI":"10.13039\/501100005788","name":"Medical University of Vienna","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100005788","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Parasitol Res"],"published-print":{"date-parts":[[2024,2]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The parasite <jats:italic>Entamoeba histolytica<\/jats:italic> is the cause of amoebic dysentery and liver abscess in humans. On the protozoan cell surface, a variety of glycosylated molecules are involved in the interaction with the environment, such as attachment to the colonic mucus. One of these molecules is the lipopeptidophosphoglycan (LPPG), a complex surface component with antigenic properties. Its structure is only partly known, it is a glycosylphosphatidylinositol (GPI)-linked glycoprotein with a large amount of O-glycosylation. To date, the sequence of a core protein has not been identified. In this study, we further investigated this complex surface molecule aided by the availability of the monoclonal antibody EH5, which had been raised in our laboratory. We studied the extraction of LPPG in various solvent mixtures and discovered that 2-butanol saturated water was simple and superior to other solvents used in the past. The isolated LPPG was subjected to treatment with several proteases and the Ser\/Thr specific cleavage agent scandium (III) trifluoromethanesulfonate (scandium triflate). The products were probed with antibody EH5 and the blots showed that the LPPG preparation was largely resistant to standard proteases, but could be cleaved by the scandium compound. These observations could point to the existence of a Ser- or Thr-rich core protein structure.<\/jats:p>","DOI":"10.1007\/s00436-024-08149-6","type":"journal-article","created":{"date-parts":[[2024,2,21]],"date-time":"2024-02-21T03:02:06Z","timestamp":1708484526000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Revisiting the isolation and characterisation of Entamoeba histolytica lipopeptidophosphoglycan"],"prefix":"10.1007","volume":"123","author":[{"given":"Anna","family":"Nagode","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jorick","family":"Vanbeselaere","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael","family":"Duch\u00eane","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,2,21]]},"reference":[{"key":"8149_CR1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0158979","volume":"11","author":"EE \u00c1vila","year":"2016","unstructured":"\u00c1vila EE, Salaiza N, Pulido J, Rodr\u00edguez MC, D\u00edaz-God\u00ednez C, Laclette JP, Becker I, Carrero JC (2016) Entamoeba histolytica trophozoites and lipopeptidophosphoglycan trigger human neutrophil extracellular traps. PLoS ONE 11:e0158979. https:\/\/doi.org\/10.1371\/journal.pone.0158979","journal-title":"PLoS ONE"},{"key":"8149_CR2","doi-asserted-by":"publisher","first-page":"1218","DOI":"10.1016\/j.vaccine.2013.06.056","volume":"32","author":"L Barroso","year":"2014","unstructured":"Barroso L, Abhyankar M, Noor Z, Read K, Pedersen K, White R, Fox C, Petri WA, Lyerly D (2014) Expression, purification, and evaluation of recombinant LecA as a candidate for an amebic colitis vaccine. Vaccine 32:1218\u20131224. https:\/\/doi.org\/10.1016\/j.vaccine.2013.06.056","journal-title":"Vaccine"},{"key":"8149_CR3","doi-asserted-by":"publisher","first-page":"1711","DOI":"10.1128\/iai.60.4.1711-1713.1992","volume":"60","author":"A Bhattacharya","year":"1992","unstructured":"Bhattacharya A, Ghildyal R, Prasad J, Bhattacharya S, Diamond LS (1992a) Modulation of a surface antigen of Entamoeba histolytica in response to bacteria. Infect Immun 60:1711\u20131713. https:\/\/doi.org\/10.1128\/iai.60.4.1711-1713.1992","journal-title":"Infect Immun"},{"key":"8149_CR4","doi-asserted-by":"publisher","first-page":"161","DOI":"10.1016\/0166-6851(92)90163-E","volume":"56","author":"A Bhattacharya","year":"1992","unstructured":"Bhattacharya A, Prasad R, Sacks DL (1992b) Identification and partial characterization of a lipophosphoglycan from a pathogenic strain of Entamoeba histolytica. Mol Biochem Parasitol 56:161\u2013168. https:\/\/doi.org\/10.1016\/0166-6851(92)90163-E","journal-title":"Mol Biochem Parasitol"},{"key":"8149_CR5","doi-asserted-by":"publisher","first-page":"31","DOI":"10.1017\/S0031182099005259","volume":"120","author":"A Bhattacharya","year":"2000","unstructured":"Bhattacharya A, Arya R, Clark CG, Ackers JP (2000) Absence of lipophosphoglycan-like glycoconjugates in Entamoeba dispar. Parasitology 120:31\u201335. https:\/\/doi.org\/10.1017\/S0031182099005259","journal-title":"Parasitology"},{"key":"8149_CR6","doi-asserted-by":"publisher","first-page":"911","DOI":"10.1139\/o59-099","volume":"37","author":"EG Bligh","year":"1959","unstructured":"Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37:911\u2013917. https:\/\/doi.org\/10.1139\/o59-099","journal-title":"Can J Biochem Physiol"},{"key":"8149_CR7","doi-asserted-by":"publisher","first-page":"9472","DOI":"10.1038\/s41598-017-09894-8","volume":"7","author":"SL Choy","year":"2017","unstructured":"Choy SL, Bernin H, Aiba T et al (2017) Synthetic analogs of an Entamoeba histolytica glycolipid designed to combat intracellular Leishmania infection. Sci Rep 7:9472. https:\/\/doi.org\/10.1038\/s41598-017-09894-8","journal-title":"Sci Rep"},{"key":"8149_CR8","doi-asserted-by":"publisher","first-page":"431","DOI":"10.1016\/0035-9203(78)90144-X","volume":"72","author":"LS Diamond","year":"1978","unstructured":"Diamond LS, Harlow DR, Cunnick CC (1978) A new medium for the axenic cultivation of Entamoeba histolytica and other Entamoeba. Trans R Soc Trop Med Hyg 72:431\u2013432. https:\/\/doi.org\/10.1016\/0035-9203(78)90144-X","journal-title":"Trans R Soc Trop Med Hyg"},{"key":"8149_CR9","doi-asserted-by":"publisher","first-page":"2088","DOI":"10.1021\/bi00859a028","volume":"6","author":"J Feder","year":"1967","unstructured":"Feder J (1967) Studies on the specificity of Bacillus subtilis neutral protease with synthetic substrates. Biochemistry 6:2088\u20132093. https:\/\/doi.org\/10.1021\/bi00859a028","journal-title":"Biochemistry"},{"key":"8149_CR10","doi-asserted-by":"publisher","first-page":"e00161","DOI":"10.1128\/AAC.00161-20","volume":"64","author":"H Fehling","year":"2020","unstructured":"Fehling H, Choy SL, Ting F, Landschulze D, Bernin H, Lender SC, M\u00fchlenpfordt M, Bifeld E, Eick J, Marggraff C, Kottmayr N, Groneberg M, Hoenow S, Sellau J, Clos J, Meier C, Lotter H (2020) Antileishmanial effects of synthetic EhPIb analogs derived from the Entamoeba histolytica lipopeptidephosphoglycan. Antimicrob Agents Chemother 64:e00161-e220. https:\/\/doi.org\/10.1128\/AAC.00161-20","journal-title":"Antimicrob Agents Chemother"},{"key":"8149_CR11","doi-asserted-by":"publisher","first-page":"53R","DOI":"10.1093\/glycob\/cwj007","volume":"15","author":"JR Frederick","year":"2005","unstructured":"Frederick JR, Petri WA (2005) Roles for the galactose-\/N-acetylgalactosamine-binding lectin of Entamoeba in parasite virulence and differentiation. Glycobiology 15:53R-59R. https:\/\/doi.org\/10.1093\/glycob\/cwj007","journal-title":"Glycobiology"},{"issue":"Suppl 3","key":"8149_CR12","first-page":"51","volume":"13","author":"A Isibasi","year":"1982","unstructured":"Isibasi A, Santa Cruz M, Ram\u00edrez A, Kumate J (1982a) Immunochemistry of a lipopeptidophosphoglycan extracted from trophozoites of Entamoeba histolytica strain HK-9 cultivated in axenic media, using the phenol-water method. Arch Invest Med (mex) 13(Suppl 3):51\u201355","journal-title":"Arch Invest Med (mex)"},{"issue":"Suppl 3","key":"8149_CR13","first-page":"57","volume":"13","author":"A Isibasi","year":"1982","unstructured":"Isibasi A, Santa Cruz M, Soto Montano X, Ram\u00edrez A, Kumate J (1982b) Localization of a lipopeptidophosphoglycan extracted by phenol-water from trophozoites of the HK-9 strain of Entamoeba histolytica. Arch Invest Med (mex) 13(Suppl 3):57\u201362","journal-title":"Arch Invest Med (mex)"},{"key":"8149_CR14","doi-asserted-by":"publisher","first-page":"61","DOI":"10.1007\/s00775-019-01733-7","volume":"25","author":"CJ Koehler","year":"2020","unstructured":"Koehler CJ, Thiede B (2020) Predominant cleavage of proteins N-terminal to serines and threonines using scandium(III) triflate. J Biol Inorg Chem 25:61\u201366. https:\/\/doi.org\/10.1007\/s00775-019-01733-7","journal-title":"J Biol Inorg Chem"},{"key":"8149_CR15","doi-asserted-by":"publisher","first-page":"3335","DOI":"10.1016\/S0021-9258(19)42577-5","volume":"249","author":"ED Korn","year":"1974","unstructured":"Korn ED, Dearborn DG, Wright PL (1974) Lipophosphonoglycan of the plasma membrance of Acanthamoeba castellanii. Isolation from whole amoebae and identification of the water-soluble products of acid hydrolysis. J Biol Chem 249:3335\u20133341","journal-title":"J Biol Chem"},{"key":"8149_CR16","doi-asserted-by":"publisher","first-page":"713","DOI":"10.1111\/j.1432-1033.2003.03975.x","volume":"271","author":"AB Kumar","year":"2004","unstructured":"Kumar AB, Gowda LR, Tharanathan RN (2004) Non-specific depolymerization of chitosan by pronase and characterization of the resultant products. Eur J Biochem 271:713\u2013723. https:\/\/doi.org\/10.1111\/j.1432-1033.2003.03975.x","journal-title":"Eur J Biochem"},{"key":"8149_CR17","doi-asserted-by":"publisher","first-page":"118","DOI":"10.1080\/19490976.2019.1614363","volume":"11","author":"A Leon-Coria","year":"2020","unstructured":"Leon-Coria A, Kumar M, Chadee K (2020) The delicate balance between Entamoeba histolytica, mucus and microbiota. Gut Microbes 11:118\u2013125. https:\/\/doi.org\/10.1080\/19490976.2019.1614363","journal-title":"Gut Microbes"},{"key":"8149_CR18","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1000434","volume":"5","author":"H Lotter","year":"2009","unstructured":"Lotter H, Gonz\u00e1lez-Rold\u00e1n N, Lindner B, Winau F, Isibasi A, Moreno-Lafont M, Ulmer AJ, Holst O, Tannich E, Jacobs T (2009) Natural killer T cells activated by a lipopeptidophosphoglycan from Entamoeba histolytica are critically important to control amebic liver abscess. PLoS Pathog 5:e1000434. https:\/\/doi.org\/10.1371\/journal.ppat.1000434","journal-title":"PLoS Pathog"},{"key":"8149_CR19","doi-asserted-by":"publisher","first-page":"1557","DOI":"10.1084\/jem.186.9.1557","volume":"186","author":"A Marinets","year":"1997","unstructured":"Marinets A, Zhang T, Guill\u00e9n N, Gounon P, Bohle B, Vollmann U, Scheiner O, Wiedermann G, Stanley SL, Duch\u00eane M (1997) Protection against invasive amebiasis by a single monoclonal antibody directed against a lipophosphoglycan antigen localized on the surface of Entamoeba histolytica. J Exp Med 186:1557\u20131565. https:\/\/doi.org\/10.1084\/jem.186.9.1557","journal-title":"J Exp Med"},{"key":"8149_CR20","doi-asserted-by":"publisher","first-page":"321","DOI":"10.1046\/j.1365-3024.2002.00463.x","volume":"24","author":"H Melzer","year":"2002","unstructured":"Melzer H, Fortugno P, Mansouri E, Felici F, Marinets A, Wiedermann G, Kollaritsch H, von Specht B-U, Duchene M (2002) Antigenicity and immunogenicity of phage library-selected peptide mimics of the major surface proteophosphoglycan antigens of Entamoeba histolytica. Parasite Immunol 24:321\u2013328. https:\/\/doi.org\/10.1046\/j.1365-3024.2002.00463.x","journal-title":"Parasite Immunol"},{"key":"8149_CR21","doi-asserted-by":"publisher","first-page":"409","DOI":"10.1006\/jmbi.2000.3577","volume":"297","author":"S Moody-Haupt","year":"2000","unstructured":"Moody-Haupt S, Patterson JH, Mirelman D, McConville MJ (2000) The major surface antigens of Entamoeba histolytica trophozoites are GPI-anchored proteophosphoglycans. J Mol Biol 297:409\u2013420. https:\/\/doi.org\/10.1006\/jmbi.2000.3577","journal-title":"J Mol Biol"},{"key":"8149_CR22","doi-asserted-by":"publisher","first-page":"427","DOI":"10.1093\/oxfordjournals.jbchem.a128914","volume":"64","author":"Y Narahashi","year":"1968","unstructured":"Narahashi Y, Shibuya K, Yanagita M (1968) Studies on proteolytic enzymes (pronase) of Streptomyces griseus K-1. J Biochem 64:427\u2013437. https:\/\/doi.org\/10.1093\/oxfordjournals.jbchem.a128914","journal-title":"J Biochem"},{"key":"8149_CR23","doi-asserted-by":"publisher","first-page":"587","DOI":"10.1271\/bbb.70537","volume":"72","author":"M Nishinoaki","year":"2008","unstructured":"Nishinoaki M, Asakura T, Watanabe T, Kunizaki E, Matsumoto M, Eto W, Tamura T, Minami M, Obata A, Abe K, Funaki J (2008) Application of an Aspergillus saitoi protease preparation to soybean curd to modify its functional and rheological properties. Biosci Biotechnol Biochem 72:587\u2013590. https:\/\/doi.org\/10.1271\/bbb.70537","journal-title":"Biosci Biotechnol Biochem"},{"key":"8149_CR24","doi-asserted-by":"publisher","first-page":"279","DOI":"10.1016\/0166-6851(92)90177-L","volume":"56","author":"R Prasad","year":"1992","unstructured":"Prasad R, Tola M, Bhattacharya S, Sharma MP, Bhattacharya A (1992) Recognition of Entamoeba histolytica lipophosphoglycan by a strain-specific monoclonal antibody and human immune sera. Mol Biochem Parasitol 56:279\u2013287. https:\/\/doi.org\/10.1016\/0166-6851(92)90177-L","journal-title":"Mol Biochem Parasitol"},{"key":"8149_CR25","doi-asserted-by":"publisher","first-page":"300","DOI":"10.1021\/pr0705035","volume":"7","author":"J Rodriguez","year":"2008","unstructured":"Rodriguez J, Gupta N, Smith RD, Pevzner PA (2008) Does trypsin cut before proline? J Proteome Res 7:300\u2013305. https:\/\/doi.org\/10.1021\/pr0705035","journal-title":"J Proteome Res"},{"key":"8149_CR26","doi-asserted-by":"publisher","first-page":"485","DOI":"10.1007\/978-1-60761-913-0_29","volume":"681","author":"SM Smith","year":"2011","unstructured":"Smith SM (2011) Strategies for the purification of membrane proteins. Methods Mol Biol 681:485\u2013496. https:\/\/doi.org\/10.1007\/978-1-60761-913-0_29","journal-title":"Methods Mol Biol"},{"key":"8149_CR27","doi-asserted-by":"publisher","first-page":"617","DOI":"10.1111\/j.1550-7408.1995.tb05916.x","volume":"42","author":"G Srivastava","year":"1995","unstructured":"Srivastava G, Anand MT, Bhattacharya S, Bhattacharya A (1995) Lipophosphoglycan is present in distinctly different form in different Entamoeba histolytica strains and absent in Entamoeba moshkovskii and Entamoeba invadens. J Eukaryot Microbiol 42:617\u2013622. https:\/\/doi.org\/10.1111\/j.1550-7408.1995.tb05916.x","journal-title":"J Eukaryot Microbiol"},{"key":"8149_CR28","doi-asserted-by":"publisher","first-page":"127","DOI":"10.1016\/0166-6851(92)90250-N","volume":"50","author":"SL Stanley","year":"1992","unstructured":"Stanley SL, Huizenga H, Li E (1992) Isolation and partial characterization of a surface glycoconjugate of Entamoeba histolytica. Mol Biochem Parasitol 50:127\u2013138. https:\/\/doi.org\/10.1016\/0166-6851(92)90250-N","journal-title":"Mol Biochem Parasitol"},{"key":"8149_CR29","doi-asserted-by":"publisher","first-page":"3883","DOI":"10.1016\/s0021-9258(17)43180-2","volume":"259","author":"SJ Turco","year":"1984","unstructured":"Turco SJ, Wilkerson MA, Clawson DR (1984) Expression of an unusual acidic glycoconjugate in Leishmania donovani. J Biol Chem 259:3883\u20133889. https:\/\/doi.org\/10.1016\/s0021-9258(17)43180-2","journal-title":"J Biol Chem"},{"key":"8149_CR30","doi-asserted-by":"publisher","first-page":"1693","DOI":"10.1038\/s41467-021-21754-8","volume":"12","author":"F Uliana","year":"2021","unstructured":"Uliana F, Vizovi\u0161ek M, Acquasaliente L, Ciuffa R, Fossati A, Frommelt F, Goetze S, Wollscheid B, Gstaiger M, De Filippis V, auf dem Keller U, Aebersold R, (2021) Mapping specificity, cleavage entropy, allosteric changes and substrates of blood proteases in a high-throughput screen. Nat Commun 12:1693. https:\/\/doi.org\/10.1038\/s41467-021-21754-8","journal-title":"Nat Commun"},{"key":"8149_CR31","doi-asserted-by":"publisher","first-page":"2069","DOI":"10.1091\/mbc.9.8.2069","volume":"9","author":"RR Vines","year":"1998","unstructured":"Vines RR, Ramakrishnan G, Rogers JB, Lockhart LA, Mann BJ, Petri WA (1998) Regulation of adherence and virulence by the Entamoeba histolytica lectin cytoplasmic domain, which contains a \u03b22 integrin motif. Mol Biol Cell 9:2069\u20132079. https:\/\/doi.org\/10.1091\/mbc.9.8.2069","journal-title":"Mol Biol Cell"},{"key":"8149_CR32","doi-asserted-by":"publisher","first-page":"7955","DOI":"10.1016\/S0021-9258(17)39545-5","volume":"260","author":"A Wesley","year":"1985","unstructured":"Wesley A, Mantle M, Man D, Qureshi R, Forstner G, Forstner J (1985) Neutral and acidic species of human intestinal mucin. Evidence for different core peptides. J Biol Chem 260:7955\u20137959. https:\/\/doi.org\/10.1016\/S0021-9258(17)39545-5","journal-title":"J Biol Chem"},{"key":"8149_CR33","unstructured":"Westphal O, Jann K (1965) Bacterial lipopolysaccharides extraction with phenol-water and further applications of the procedure. Methods Carbohydr Chem 5:83\u201391"},{"key":"8149_CR34","doi-asserted-by":"publisher","first-page":"5873","DOI":"10.1128\/IAI.70.10.5873-5876.2002","volume":"70","author":"Z Zhang","year":"2002","unstructured":"Zhang Z, Duch\u00eane M, Stanley SL (2002) A monoclonal antibody to the amebic lipophosphoglycan-proteophosphoglycan antigens can prevent disease in human intestinal xenografts infected with Entamoeba histolytica. Infect Immun 70:5873\u20135876. https:\/\/doi.org\/10.1128\/IAI.70.10.5873-5876.2002","journal-title":"Infect Immun"}],"container-title":["Parasitology Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s00436-024-08149-6.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1007\/s00436-024-08149-6\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s00436-024-08149-6.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,3,8]],"date-time":"2024-03-08T21:50:39Z","timestamp":1709934639000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1007\/s00436-024-08149-6"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,2]]},"references-count":34,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2024,2]]}},"alternative-id":["8149"],"URL":"https:\/\/doi.org\/10.1007\/s00436-024-08149-6","relation":{},"ISSN":["0932-0113","1432-1955"],"issn-type":[{"value":"0932-0113","type":"print"},{"value":"1432-1955","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,2]]},"assertion":[{"value":"21 November 2023","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"31 January 2024","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"21 February 2024","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The authors declare no competing interests.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"138"}}