Nanobody Mediated Macromolecular Crowding Induces Membrane Fission and Remodeling in the African Trypanosome

Hempelmann, A.; Hartleb, L.; Van Straaten, M.; Hashemi, H.; Zeelen, J. P.; Papavasiliou, F. N.; Engstler, M.; Stebbins, C. E.; Jones, N. G.

doi:10.1101/2021.01.13.426364

bioRxiv · 10.1101/2021.01.13.426364

Nanobody Mediated Macromolecular Crowding Induces Membrane Fission and Remodeling in the African Trypanosome

Hempelmann, A.·Hartleb, L.·Van Straaten, M.·Hashemi, H.·Zeelen, J. P.·Papavasiliou, F. N.·Engstler, M.·Stebbins, C. E.·Jones, N. G.

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Abstract

The dense Variant Surface Glycoprotein (VSG) coat of African trypanosomes represents the primary host-pathogen interface. Antigenic variation prevents clearing of the pathogen by employing a large repertoire of antigenically distinct VSG genes, thus neutralizing the hosts antibody response. To explore the epitope space of VSGs, we generated anti-VSG nanobodies and combined high-resolution structural analysis of VSG-nanobody complexes with binding assays on living cells, revealing that these camelid antibodies bind deeply inside the coat. One nanobody caused rapid loss of cellular motility, possibly due to blockage of VSG mobility on the coat, whose rapid endo-and exocytosis is mechanistically linked to T. brucei propulsion and whose density is required for survival. Electron microscopy studies demonstrated that this loss of motility was accompanied by rapid formation and shedding of nanovesicles and nanotubes, suggesting that increased protein crowding on the dense membrane can be a driving force for membrane fission in living cells.

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Hempelmann, A., Hartleb, L., Van Straaten, M., Hashemi, H., Zeelen, J. P., Papavasiliou, F. N., Engstler, M., Stebbins, C. E., Jones, N. G.. 2021-01-14. Nanobody Mediated Macromolecular Crowding Induces Membrane Fission and Remodeling in the African Trypanosome. https://doi.org/10.1101/2021.01.13.426364

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