Biochemical, Conformational, and Immunogenic Analysis of Soluble Trimeric Forms of Henipavirus Fusion Glycoproteins

• DOI: 10.1128/jvi.01318-12
• Publication Date: 2012-08-23T00:17:20Z
• Authors: Yee-Peng Chan; Min Lu; Somnath Dutta; Lianying Yan; Jennifer Barr; Michael Flora; Yan-Ru Feng; Kai Xu; Dimitar B. Nikolov; Lin-Fa Wang; Georgios Skiniotis; Christopher C. Br...
• Keywords: Hendra Virus; Furin; Heptad repeat

ABSTRACT The henipaviruses, Hendra virus (HeV) and Nipah (NiV), are paramyxoviruses discovered in the mid- to late 1990s that possess a broad host tropism known cause severe often fatal disease both humans animals. HeV NiV infect cells by pH-independent membrane fusion mechanism facilitated their attachment (G) (F) glycoproteins. Here, several soluble forms of henipavirus F (sF) were engineered characterized. Recombinant sF was produced deleting transmembrane (TM) cytoplasmic tail (CT) domains appending glycosylphosphatidylinositol (GPI) anchor signal sequence followed GPI-phospholipase D digestion, trimeric coiled-coil (GCNt) domain (sF GCNt ), or TM, CT, peptide domain. These glycoproteins as 0 precursors, all apparent stable trimers recognized NiV-specific antisera. Surprisingly, however, only GCNt-appended constructs ) could elicit cross-reactive henipavirus-neutralizing antibody mice. In addition, be triggered vitro protease cleavage heat transition from an prefusion postfusion conformation, transitioning through intermediate captured corresponding C-terminal heptad repeat F. pre- structures non-GCNt-appended revealed electron microscopy distinguishable F-specific monoclonal antibodies. data suggest certain serve potential subunit vaccine immunogens against henipaviruses also establish important tools for further structural, functional, diagnostic studies on these emerging viruses.