{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,11]],"date-time":"2025-11-11T22:18:27Z","timestamp":1762899507748},"reference-count":60,"publisher":"Springer Science and Business Media LLC","issue":"1","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2018,3,26]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Following the outcome from a previously performed yeast two-hybrid experiment, the binding strength between<jats:italic>T<\/jats:italic>.<jats:italic>gondii<\/jats:italic>SAG1 and SAG2 and their respective prey proteins were further confirmed in this study. The<jats:italic>sag1<\/jats:italic>,<jats:italic>sag2<\/jats:italic>and their prey genes were amplified and cloned into a pGEMT vector. To express the recombinant proteins, the fragments were then subcloned into a pRSETA vector and transformed into<jats:italic>E<\/jats:italic>.<jats:italic>coli<\/jats:italic>BL21 (DE3) cells. The recombinant proteins were expressed optimally at 37\u00b0C and 1mM of IPTG. The 6X His-tag fusion proteins were purified, dialyzed and concentrated. To confirm the expressed proteins, the recombinant proteins were analysed by SDS-PAGE and Western blot. As expected, the size of SAG1, SAG2, HLY and HZF protein were 32, 23, 28 and 37 kDa, respectively. The purified proteins were loaded onto a MicroCal Auto-iTC<jats:sub>200<\/jats:sub>calorimeter from MicroCal\u2122 to quantify binding strength. ITC results indicated there was a typical binding curve for interactions between SAG1 and HLY protein. However, there was an atypical binding curve obtained for interactions between SAG2 and HZF protein. By observing the data obtained from the ITC assay, both of the human proteins (HLY and HZF) were demonstrated to bind to their respective SAG1 and SAG2 proteins.<\/jats:p>","DOI":"10.1515\/ap-2018-0012","type":"journal-article","created":{"date-parts":[[2018,1,19]],"date-time":"2018-01-19T22:15:48Z","timestamp":1516400148000},"page":"106-113","source":"Crossref","is-referenced-by-count":6,"title":["Measurement of binding strength between prey proteins interacting with Toxoplasma gondii SAG1 and SAG2 using isothermal titration calorimetry (ITC)"],"prefix":"10.1007","volume":"63","author":[{"given":"Meng-Yee","family":"Lai","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yee-Ling","family":"Lau","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","reference":[{"key":"ref31","doi-asserted-by":"crossref","first-page":"8162","DOI":"10.1016\/S0021-9258(17)34376-4","article-title":"Protein associations and basic protein conformation in the myelin membrane","volume":"253","year":"1978","journal-title":"The Journal of Biological Chemistry"},{"key":"ref241","doi-asserted-by":"crossref","first-page":"6680","DOI":"10.1128\/IAI.73.10.6680-6688.2005","article-title":"Disruption of a locus encoding a nucleolar zinc finger protein decreases tachyzoite-to-radyzoite differentiation in Toxoplasma gondii","volume":"73","year":"2005","journal-title":"Infection and Immunology"},{"key":"ref451","first-page":"351","article-title":"Attachment of Toxoplasma gondii to host cells involves major surface protein, SAG1-1 (P30)","volume":"79","year":"1994","journal-title":"Experimental Parasitology"},{"key":"ref151","first-page":"351","article-title":"Attachment of Toxoplasma gondii to host cells involves major surface protein, SAG1-1 (P30)","volume":"79","year":"1994","journal-title":"Experimental Parasitology"},{"key":"ref351","article-title":"The motility of a human parasite, Toxoplasma gondii, is regulated by a novel lysine methyltransferase","volume":"7","year":"2011","journal-title":"PLOS Pathogen"},{"key":"ref101","doi-asserted-by":"crossref","first-page":"560","DOI":"10.1016\/S0959-440X(00)00248-7","article-title":"Direct measurement of protein binding energetics by isothermal titration calorimetry","volume":"11","year":"2001","journal-title":"Current Opinion in Structural Biology"},{"key":"ref331","doi-asserted-by":"crossref","first-page":"8162","DOI":"10.1016\/S0021-9258(17)34376-4","article-title":"Protein associations and basic protein conformation in the myelin membrane","volume":"253","year":"1978","journal-title":"The Journal of Biological Chemistry"},{"key":"ref481","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/j.bbamem.2013.05.023","article-title":"Isothermal titration calorimetry of membrane proteins-progress and challenges","volume":"1838","year":"2014","journal-title":"Biochemica et Biophysica Acta"},{"key":"ref541","doi-asserted-by":"crossref","first-page":"6680","DOI":"10.1128\/IAI.73.10.6680-6688.2005","article-title":"Disruption of a locus encoding a nucleolar zinc finger protein decreases tachyzoite-to-radyzoite differentiation in Toxoplasma gondii","volume":"73","year":"2005","journal-title":"Infection and Immunology"},{"key":"ref531","doi-asserted-by":"crossref","first-page":"3199","DOI":"10.1038\/srep03199","article-title":"Characterization of the interaction between Toxoplasma gondii rhoptry neck protein 4 and host cellular \u03b2-tubulin","volume":"3","year":"2013","journal-title":"Scientific Reports"},{"key":"ref21","first-page":"244","article-title":"Production of recombinant proteins from protozoan parasites","volume":"5","year":"2010","journal-title":"Trends Parasitology"},{"key":"ref511","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1038\/359082a0","article-title":"Virulent strains of Toxoplasma gondii comprise a single clonal lineage","volume":"359","year":"1992","journal-title":"Nature"},{"key":"ref411","first-page":"1285","article-title":"Surface antigens of Toxoplasma gondii: Variations on a theme","volume":"31","year":"2000","journal-title":"International Journal of Parasitology"},{"key":"ref501","first-page":"452","article-title":"Yeast two-hybrid and ITC studies of alpha and beta spectrin interaction at the tetramerization site","volume":"16","year":"2001","journal-title":"Cell Molecular Biology Letter"},{"key":"ref341","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1016\/0020-7519(95)00103-4","article-title":"Toxoplasma gondii: The role of parasite surface and secreted proteins in host cell invasion","volume":"26","year":"1996","journal-title":"International Journal of Parasitology"},{"key":"ref561","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1016\/j.vetpar.2008.03.032","article-title":"High-level expression of SAG1 and GRA7 gene of Toxoplasma gondii (Izatnagar isolate) and their application in serodiagnosis of goat toxoplasmosis","volume":"154","year":"2008","journal-title":"Veterinay Parasitology"},{"key":"ref591","first-page":"412","article-title":"Efficient soluble expression, purification and identification of the truncated SAG1 gene of Toxoplasma gondii in Escherichia coli","volume":"4","year":"2004","journal-title":"Journal of First Military Medical University"},{"key":"ref381","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1016\/S0959-440X(00)00167-6","article-title":"Zinc finger proteins: new insights into structural and functional diversity","volume":"11","year":"2001","journal-title":"Current Opinion Structural Biology"},{"key":"ref201","first-page":"452","article-title":"Yeast two-hybrid and ITC studies of alpha and beta spectrin interaction at the tetramerization site","volume":"16","year":"2001","journal-title":"Cell Molecular Biology Letter"},{"key":"ref401","doi-asserted-by":"crossref","first-page":"560","DOI":"10.1016\/S0959-440X(00)00248-7","article-title":"Direct measurement of protein binding energetics by isothermal titration calorimetry","volume":"11","year":"2001","journal-title":"Current Opinion in Structural Biology"},{"key":"ref111","first-page":"1285","article-title":"Surface antigens of Toxoplasma gondii: Variations on a theme","volume":"31","year":"2000","journal-title":"International Journal of Parasitology"},{"key":"ref181","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/j.bbamem.2013.05.023","article-title":"Isothermal titration calorimetry of membrane proteins-progress and challenges","volume":"1838","year":"2014","journal-title":"Biochemica et Biophysica Acta"},{"key":"ref221","doi-asserted-by":"crossref","first-page":"414","DOI":"10.1126\/science.1078035","article-title":"Recent expansion of Toxoplasma through enhanced oral transmission","volume":"299","year":"2003","journal-title":"Science"},{"key":"ref161","article-title":"Antibodies to Toxoplasma gondii major surface protein (SAG-1, P30) inhibit infection of host cells and are produced in mapical membrane antigen 1 (AMA1) governs ligand binding selectivity","volume":"10","year":"1993","journal-title":"Plos One"},{"key":"ref461","article-title":"Antibodies to Toxoplasma gondii major surface protein (SAG-1, P30) inhibit infection of host cells and are produced in mapical membrane antigen 1 (AMA1) governs ligand binding selectivity","volume":"10","year":"1993","journal-title":"Plos One"},{"key":"ref441","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s13071-016-1943-1","article-title":"Identification of host proteins interacting with Toxoplasma gondii GRA15 (TgGRA15) by yeast two-hybrid system","volume":"10","year":"2017","journal-title":"Parasites & Vectors"},{"key":"ref41","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1016\/0020-7519(95)00103-4","article-title":"Toxoplasma gondii: The role of parasite surface and secreted proteins in host cell invasion","volume":"26","year":"1996","journal-title":"International Journal of Parasitology"},{"key":"ref191","doi-asserted-by":"crossref","first-page":"391","DOI":"10.1017\/S0031182003004736","article-title":"Toxoplasma gondii major surface antigen (SAG1): in vitro analysis of host cell binding","volume":"128","year":"2004","journal-title":"Parasitology"},{"key":"ref121","doi-asserted-by":"crossref","first-page":"256","DOI":"10.1016\/0167-4838(88)90142-2","article-title":"Catalysis of proline isomerization during protein-folding reactions","volume":"956","year":"1988","journal-title":"Biochimica et Biophysica Acta"},{"key":"ref131","doi-asserted-by":"crossref","first-page":"4028","DOI":"10.1073\/pnas.87.11.4028","article-title":"Peptidyl-prolyl cis-trans-isomerase from Escherichia coli: a periplasmic homolog of cyclophilin that is not inhibited by cyclosporin A","volume":"87","year":"1990","journal-title":"Proceedings of the National Academy Sciences United States America"},{"key":"ref571","doi-asserted-by":"crossref","first-page":"180","DOI":"10.1186\/1756-3305-7-180","article-title":"Research progress on surface antigen 1 (SAG1) of Toxoplasma gondii","volume":"7","year":"2014","journal-title":"Parasites & Vectors"},{"key":"ref141","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s13071-016-1943-1","article-title":"Identification of host proteins interacting with Toxoplasma gondii GRA15 (TgGRA15) by yeast two-hybrid system","volume":"10","year":"2017","journal-title":"Parasites & Vectors"},{"key":"ref51","article-title":"The motility of a human parasite, Toxoplasma gondii, is regulated by a novel lysine methyltransferase","volume":"7","year":"2011","journal-title":"PLOS Pathogen"},{"key":"ref91","first-page":"1900","article-title":"Host cell manipulation by the human pathogen Toxoplasma gondii","volume":"5","year":"2008","journal-title":"Cell Molecular Life Sciences"},{"key":"ref301","doi-asserted-by":"crossref","first-page":"303","DOI":"10.3347\/kjp.2006.44.4.303","article-title":"Interactions between secreted GRA proteins and host cell proteins across the parasitophorous vacuolar membrane in the parasitism of Toxoplasma gondii","volume":"44","year":"2006","journal-title":"Korean Journal of Parasitology"},{"key":"ref251","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1017\/S0031182001008460","article-title":"Anti-SAG1peptide antibodies inhibit the penetration of Toxoplasma gondii tachyzoites into enterocyte cell lines","volume":"123","year":"2001","journal-title":"Parasitology"},{"key":"ref11","doi-asserted-by":"crossref","first-page":"7221","DOI":"10.1073\/pnas.94.14.7221","article-title":"Slow dimer dissociation of the TATA binding protein dictates the kinetics of DNA binding","volume":"94","year":"1997","journal-title":"Proceedings of the National Academy Sciences United States America"},{"key":"ref01","doi-asserted-by":"crossref","first-page":"303","DOI":"10.3347\/kjp.2006.44.4.303","article-title":"Interactions between secreted GRA proteins and host cell proteins across the parasitophorous vacuolar membrane in the parasitism of Toxoplasma gondii","volume":"44","year":"2006","journal-title":"Korean Journal of Parasitology"},{"key":"ref71","doi-asserted-by":"crossref","first-page":"456","DOI":"10.1186\/s13071-017-2387-y","article-title":"Screening and identification of host proteins interacting with Toxoplasma gondii SAG2 by yeast two-hybrid assay","volume":"10","year":"2017","journal-title":"Parasites & Vectors"},{"key":"ref231","doi-asserted-by":"crossref","first-page":"3199","DOI":"10.1038\/srep03199","article-title":"Characterization of the interaction between Toxoplasma gondii rhoptry neck protein 4 and host cellular \u03b2-tubulin","volume":"3","year":"2013","journal-title":"Scientific Reports"},{"key":"ref321","first-page":"244","article-title":"Production of recombinant proteins from protozoan parasites","volume":"5","year":"2010","journal-title":"Trends Parasitology"},{"key":"ref551","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1017\/S0031182001008460","article-title":"Anti-SAG1peptide antibodies inhibit the penetration of Toxoplasma gondii tachyzoites into enterocyte cell lines","volume":"123","year":"2001","journal-title":"Parasitology"},{"key":"ref291","first-page":"412","article-title":"Efficient soluble expression, purification and identification of the truncated SAG1 gene of Toxoplasma gondii in Escherichia coli","volume":"4","year":"2004","journal-title":"Journal of First Military Medical University"},{"key":"ref521","doi-asserted-by":"crossref","first-page":"414","DOI":"10.1126\/science.1078035","article-title":"Recent expansion of Toxoplasma through enhanced oral transmission","volume":"299","year":"2003","journal-title":"Science"},{"key":"ref361","article-title":"Mechanisms and pathways of Toxoplasma gondii transepithelial migration","volume":"5","year":"2017","journal-title":"Tissue Barriers"},{"key":"ref271","doi-asserted-by":"crossref","first-page":"180","DOI":"10.1186\/1756-3305-7-180","article-title":"Research progress on surface antigen 1 (SAG1) of Toxoplasma gondii","volume":"7","year":"2014","journal-title":"Parasites & Vectors"},{"key":"ref371","doi-asserted-by":"crossref","first-page":"456","DOI":"10.1186\/s13071-017-2387-y","article-title":"Screening and identification of host proteins interacting with Toxoplasma gondii SAG2 by yeast two-hybrid assay","volume":"10","year":"2017","journal-title":"Parasites & Vectors"},{"key":"ref391","first-page":"1900","article-title":"Host cell manipulation by the human pathogen Toxoplasma gondii","volume":"5","year":"2008","journal-title":"Cell Molecular Life Sciences"},{"key":"ref421","doi-asserted-by":"crossref","first-page":"256","DOI":"10.1016\/0167-4838(88)90142-2","article-title":"Catalysis of proline isomerization during protein-folding reactions","volume":"956","year":"1988","journal-title":"Biochimica et Biophysica Acta"},{"key":"ref311","doi-asserted-by":"crossref","first-page":"7221","DOI":"10.1073\/pnas.94.14.7221","article-title":"Slow dimer dissociation of the TATA binding protein dictates the kinetics of DNA binding","volume":"94","year":"1997","journal-title":"Proceedings of the National Academy Sciences United States America"},{"key":"ref491","doi-asserted-by":"crossref","first-page":"391","DOI":"10.1017\/S0031182003004736","article-title":"Toxoplasma gondii major surface antigen (SAG1): in vitro analysis of host cell binding","volume":"128","year":"2004","journal-title":"Parasitology"},{"key":"ref281","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/j.jsb.2011.07.011","article-title":"Solution structure and dynamics of ADF from Toxoplasma gondii","volume":"176","year":"2011","journal-title":"Journal of Structural Biology"},{"key":"ref61","article-title":"Mechanisms and pathways of Toxoplasma gondii transepithelial migration","volume":"5","year":"2017","journal-title":"Tissue Barriers"},{"key":"ref261","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1016\/j.vetpar.2008.03.032","article-title":"High-level expression of SAG1 and GRA7 gene of Toxoplasma gondii (Izatnagar isolate) and their application in serodiagnosis of goat toxoplasmosis","volume":"154","year":"2008","journal-title":"Veterinay Parasitology"},{"key":"ref581","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/j.jsb.2011.07.011","article-title":"Solution structure and dynamics of ADF from Toxoplasma gondii","volume":"176","year":"2011","journal-title":"Journal of Structural Biology"},{"key":"ref431","doi-asserted-by":"crossref","first-page":"4028","DOI":"10.1073\/pnas.87.11.4028","article-title":"Peptidyl-prolyl cis-trans-isomerase from Escherichia coli: a periplasmic homolog of cyclophilin that is not inhibited by cyclosporin A","volume":"87","year":"1990","journal-title":"Proceedings of the National Academy Sciences United States America"},{"key":"ref471","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1006\/meth.1999.0852","article-title":"Isothermal titration calorimetry of protein-protein interactions","volume":"19","year":"1999","journal-title":"Methods"},{"key":"ref81","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1016\/S0959-440X(00)00167-6","article-title":"Zinc finger proteins: new insights into structural and functional diversity","volume":"11","year":"2001","journal-title":"Current Opinion Structural Biology"},{"key":"ref171","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1006\/meth.1999.0852","article-title":"Isothermal titration calorimetry of protein-protein interactions","volume":"19","year":"1999","journal-title":"Methods"},{"key":"ref211","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1038\/359082a0","article-title":"Virulent strains of Toxoplasma gondii comprise a single clonal lineage","volume":"359","year":"1992","journal-title":"Nature"}],"container-title":["Acta Parasitologica"],"original-title":[],"link":[{"URL":"http:\/\/www.degruyter.com\/view\/j\/ap.2018.63.issue-1\/ap-2018-0012\/ap-2018-0012.xml","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"http:\/\/www.degruyter.com\/view\/j\/ap.2018.63.issue-1\/ap-2018-0012\/ap-2018-0012.xml","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,8,12]],"date-time":"2022-08-12T23:00:57Z","timestamp":1660345257000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.degruyter.com\/doi\/10.1515\/ap-2018-0012"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,3,26]]},"references-count":60,"journal-issue":{"issue":"1"},"URL":"https:\/\/doi.org\/10.1515\/ap-2018-0012","relation":{},"ISSN":["1896-1851","1230-2821"],"issn-type":[{"value":"1896-1851","type":"electronic"},{"value":"1230-2821","type":"print"}],"subject":[],"published":{"date-parts":[[2018,3,26]]}}}