| dc.contributor.advisor | Janshoff, Andreas Prof. Dr. | |
| dc.contributor.author | Bao, Chunxiao | |
| dc.date.accessioned | 2014-04-25T08:03:53Z | |
| dc.date.available | 2015-05-15T22:50:05Z | |
| dc.date.issued | 2014-04-25 | |
| dc.identifier.uri | http://hdl.handle.net/11858/00-1735-0000-0022-5E98-A | |
| dc.identifier.uri | http://dx.doi.org/10.53846/goediss-4478 | |
| dc.language.iso | eng | de |
| dc.publisher | Niedersächsische Staats- und Universitätsbibliothek Göttingen | de |
| dc.rights.uri | http://creativecommons.org/licenses/by-nc-nd/3.0/ | |
| dc.subject.ddc | 540 | de |
| dc.title | Optical Fusion Assay Based on Membrane-Coated Beads in a 2D Assembly | de |
| dc.type | doctoralThesis | de |
| dc.contributor.referee | Janshoff, Andreas Prof. Dr. | |
| dc.date.examination | 2014-04-02 | |
| dc.description.abstracteng | A novel assay of membrane fusion is established based on membrane-coated beads in a 2D assembly. This assay allows identifying the different stages of the fusion process. Membrane fusion driven by heterodimeric coiled coil formation as a proof of concept using fusogenic K- and E-peptides provided results comparable to those obtained with conventional liposome assays, but with additional information on docking efficiency.
In this fusion assay, LBs and SBs are modified with i-K3Cys (LB-i-K3) and i-E3Cys (SB-i-E3) respectively and LBs are fluorescent labeled. The populations of fusogenic beads can be distinguished by size discrimination. When LB-i-K3and SB-i-E3 are mixed in water, the fusion process terminates in hemifusion. Interestingly, all fusion events including docking, hemifusion and full fusion are observed in the presence of Ca2+ in one experiment. This may due to the bridging effect of calcium ions binding to PC and non-reacted negative MCC-DOPE.
Block or change the zipping conformational of coiled coil structure is necessary for the viral infection of host cells. In this bead-based assay, the introduction of inhibitors (i-E3Cys) efficiently decreases the hemifusion efficiency and abolishes full fusion (in the presence of Ca2+). Therefore, this approach is expected to be an invaluable tool to identify small-molecule inhibitors of viral fusion with unprecedented accuracy in prefusion state.
Multiple fusion could be observed and distinguished, which takes place among more than two fusogenic membrane-coated beads. The orientations of coiled coil formation, including parallel and antiparallel orientations, are shown to have no effect on the fusion process in this bead-based assay. Most interestingly, size-dependent assays employed beads with increasing diameter scale, demonstrate that van der Waals interaction could provided energy for supported membrane detaching from the silica beads, hence promote membrane fusion. | de |
| dc.contributor.coReferee | Vana, Philipp Prof. Dr. | |
| dc.subject.eng | membrane fusion | de |
| dc.subject.eng | membrane-coated beads | de |
| dc.identifier.urn | urn:nbn:de:gbv:7-11858/00-1735-0000-0022-5E98-A-7 | |
| dc.affiliation.institute | Fakultät für Chemie | de |
| dc.subject.gokfull | Chemie (PPN62138352X) | de |
| dc.description.embargoed | 2015-05-15 | |
| dc.identifier.ppn | 783865511 | |