PMID:26585033

Citation	Fischer, K, dos Reis, VP, Finke, S, Sauerhering, L, Stroh, E, Karger, A, Maisner, A, Groschup, MH, Diederich, S and Balkema-Buschmann, A (2016) Expression, characterisation and antigenicity of a truncated Hendra virus attachment protein expressed in the protozoan host Leishmania tarentolae. J. Virol. Methods 228:48-54
Abstract	Hendra virus (HeV) is an emerging zoonotic paramyxovirus within the genus Henipavirus that has caused severe morbidity and mortality in humans and horses in Australia since 1994. HeV infection of host cells is mediated by the membrane bound attachment (G) and fusion (F) glycoproteins, that are essential for receptor binding and fusion of viral and cellular membranes. The eukaryotic unicellular parasite Leishmania tarentolae has recently been established as a powerful tool to express recombinant proteins with mammalian-like glycosylation patterns, but only few viral proteins have been expressed in this system so far. Here, we describe the purification of a truncated, Strep-tag labelled and soluble version of the HeV attachment protein (sHeV G) expressed in stably transfected L. tarentolae cells. After Strep-tag purification the identity of sHeV G was confirmed by immunoblotting and mass spectrometry. The functional binding of sHeV G to the HeV cell entry receptor ephrin-B2 was confirmed in several binding assays. Generated polyclonal rabbit antiserum against sHeV G reacted with both HeV and Nipah virus (NiV) G proteins in immunofluorescence assay and efficiently neutralised NiV infection, thus further supporting the preserved antigenicity of the purified protein.
Links	PubMed Online version:10.1016/j.jviromet.2015.11.006
Keywords	Animals; Australia; Ephrin-B2/metabolism; Hendra Virus/chemistry; Hendra Virus/genetics; Hendra Virus/immunology; Hendra Virus/physiology; Horses; Humans; Leishmania/genetics; Leishmania/metabolism; Oligopeptides/metabolism; Protein Engineering; Rabbits; Receptors, Virus/metabolism; Recombinant Proteins/immunology; Recombinant Proteins/isolation & purification; Recombinant Proteins/metabolism; Viral Envelope Proteins/genetics; Viral Envelope Proteins/immunology; Viral Envelope Proteins/isolation & purification; Viral Envelope Proteins/metabolism; Virus Attachment; Virus Internalization
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
9MONO:G5CPX1		GO:0046813: receptor-mediated virion attachment to host cell	ECO:0000314:		P		Figure 2 shows the binding of sHeV G/attachment glycoprotein/HeV attachment protein from Heniparvirus Hendra with EFNB2/Fc protein using an ELISA plate.	complete CACAO 13332

References

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