Structural and functional characterization of Plasmodium falciparum 6-Cys proteins
| dc.contributor.author | Peng, Fangni | |
| dc.contributor.supervisor | Boulanger, Martin J. | |
| dc.date.accessioned | 2016-01-06T23:33:23Z | |
| dc.date.available | 2016-11-13T12:22:06Z | |
| dc.date.copyright | 2015 | en_US |
| dc.date.issued | 2016-01-06 | |
| dc.degree.department | Department of Biochemistry and Microbiology | |
| dc.degree.level | Master of Science M.Sc. | en_US |
| dc.description.abstract | Plasmodium falciparum is the etiological agent of severe human malaria. The virulence of the parasite is dependent on a complex life cycle supported by a diverse repertoire of stage specific surface antigens. Notably, members of the 6-Cys s48/45 protein family are differentially presented on the parasite surface of each life cycle stage and known to play important biological roles, though the underlying molecular mechanisms are not well understood. Of the 6-Cys antigens, Pf41 is localized to the surface of the blood-stage merozoite through its interaction with Pf12 and is a target of the host immune system; accordingly, Pf41 is one of the five top-ranked potential malaria vaccine candidates. Pfs47 is localized to the surface of the sexual-stage gametocyte through its glycophosphatidylinositol-anchor and is currently being investigated as a transmission blocking vaccine. Intriguingly, both Pf41 and Pfs47 are predicted to adopt a three domain architecture. Prior to the studies presented here, only a single two domain 6-Cys protein had been structurally characterized. During my graduate studies, the structure of Pf41 was also determined by Dr. Michelle Parker in the Boulanger lab and I was able to perform the structural interpretation. Structural analysis revealed an unexpected topology where domains 1 and 2 are juxtaposed and the predicted central domain, which was largely proteolyzed during the crystallization process, is inserted as an extended loop in domain 1. Data from my ITC binding studies and protease protection assays suggest this inserted domain-like region (ID) plays an essential role in promoting assembly with Pf12. Despite several attempts, I was unable to crystallize Pfs47. Thus, to obtain architectural information describing Pfs47, a chemical cross linking experiment coupled with mass spectrometry was performed. The resulting data led me to predict that Pfs47 also incorporates an ID (Ser155 to Gln267) within D1. An engineered Pfs47 construct lacking the predicted ID was purified as a monomer, indicating that the predicted ID is expendable for stability of the overall structure. Collectively, these data provide important insight into the overall architecture of the biologically important Plasmodium 6-Cys proteins, which enables us to support ongoing collaborative vaccine design efforts. | en_US |
| dc.description.scholarlevel | Graduate | en_US |
| dc.identifier.uri | http://hdl.handle.net/1828/7019 | |
| dc.language | English | eng |
| dc.language.iso | en | en_US |
| dc.rights | Available to the World Wide Web | en_US |
| dc.subject | Plasmodium falciparum | en_US |
| dc.subject | 6-Cys proteins | en_US |
| dc.subject | antigens | en_US |
| dc.title | Structural and functional characterization of Plasmodium falciparum 6-Cys proteins | en_US |
| dc.type | Thesis | en_US |