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One step involves the adhesion of<jats:italic>Plasmodium<\/jats:italic>reticulocyte binding-like (RBL) proteins.<jats:italic>Plasmodium knowlesi<\/jats:italic>merozoites express only two RBL invasion ligands, known as Normocyte Binding Proteins (PkNBPXa and PkNBPXb).<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Overlapping N-terminal regions of PkNBPXa and PkNBPXb were expressed in COS7 cells and tested for surface expression and adhesion to rhesus monkey erythrocytes. Subsequent tests to study specific receptor ligand interactions included adhesion to a panel of human and non-human primate erythrocytes, enzymatic treatment, and site directed mutagenesis.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>An N-terminal cysteine-rich region of PkNBPXb (PkNBPXb-II) exhibited specific adhesion to rhesus monkey erythrocytes. Mutation of four of five cysteines in PkNBPXb-II interfered with its surface expression on COS7 cells, suggesting disulphide bond conformation is critical for intracellular trafficking. Binding of PkNBPXb-II was abolished when rhesus erythrocytes were pre-treated with chymotrypsin, but not trypsin or neuraminidase. PkNBPXb-II also bound other Old World monkey species and gibbon erythrocytes. However, erythrocytes from other primate species including humans did not bind to PkNBPXb-II or native PkNBPXb. Importantly, unlike PkNBPXb, PkNBPXa bound human erythrocytes, and this binding was independent of the Duffy blood group determinant.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>The data reported here begins to clarify the functional domains of the<jats:italic>P. knowlesi<\/jats:italic>RBLs. A binding domain has been identified and characterized in PkNBPXb. Notably, this study demonstrates that unlike PkNBPXb, PkNBPXa can bind to human erythrocytes, suggesting that PkNBPXa may function as a ligand to enable the invasion of<jats:italic>P. knowlesi<\/jats:italic>merozoites into human cells.<\/jats:p><\/jats:sec>","DOI":"10.1186\/1475-2875-11-228","type":"journal-article","created":{"date-parts":[[2012,7,6]],"date-time":"2012-07-06T16:30:55Z","timestamp":1341592255000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Two functional reticulocyte binding-like (RBL) invasion ligands of zoonotic Plasmodium knowlesi exhibit differential adhesion to monkey and human erythrocytes"],"prefix":"10.1186","volume":"11","author":[{"given":"Amma A","family":"Semenya","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tuan M","family":"Tran","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Esmeralda VS","family":"Meyer","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"John W","family":"Barnwell","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mary R","family":"Galinski","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2012,7,6]]},"reference":[{"key":"2155_CR1","volume-title":"World Malaria Report","author":"World Health Organization","year":"2011","unstructured":"World Health Organization: World Malaria Report. 2011, World Health Organization Press, Geneva"},{"key":"2155_CR2","volume-title":"Informal consultation on the public health importance of Plasmodium knowlesi","author":"World Health Organization Regional Office for the Western Pacific","year":"2011","unstructured":"World Health Organization Regional Office for the Western Pacific: Informal consultation on the public health importance of Plasmodium knowlesi. 2011, World Health Organization Press, Kuching, Sarawak, Malaysia"},{"key":"2155_CR3","doi-asserted-by":"publisher","first-page":"1017","DOI":"10.1016\/S0140-6736(04)15836-4","volume":"363","author":"B Singh","year":"2004","unstructured":"Singh B, Kim Sung L, Matusop A, Radhakrishnan A, Shamsul SS, Cox-Singh J, Thomas A, Conway DJ: A large focus of naturally acquired Plasmodium knowlesi infections in human beings. 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