title: Synthesis and secretion of recombinant tick-borne encephalitis virus protein E in soluble and particulate form.
abstract: A quantitative study was performed to investigate the requirements for secretion of recombinant soluble and particulate forms of the envelope glycoprotein E of tick-borne encephalitis (TBE) virus. Full-length E and a carboxy terminally truncated anchor-free form were expressed in COS cells in the presence and absence of prM, the precursor of the viral membrane protein M. Formation of a heteromeric complex with prM was found to be necessary for efficient secretion of both forms of E, whereas only low levels of anchor-free E were secreted in the absence of prM. The prM-mediated transport function could also be provided by coexpression of prM and E from separate constructs, but a prM-to-E ratio of greater than 1:1 did not further enhance secretion. Full-length E formed stable intracellular heterodimers with prM and was secreted as a subviral particle, whereas anchor-free E was not associated with particles and formed a less stable complex with prM, suggesting that prM interacts with both the ectodomain and anchor region of E.
authors.affiliation: Institute of Virology, University of Vienna, Austria.
authors.forename: S L
authors.initials: SL
authors.lastname: Allison
authors.forename: K
authors.initials: K
authors.lastname: Stadler
authors.forename: C W
authors.initials: CW
authors.lastname: Mandl
authors.forename: C
authors.initials: C
authors.lastname: Kunz
authors.forename: F X
authors.initials: FX
authors.lastname: Heinz
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D002460:Cell Line; D002522:Chlorocebus aethiops; D004591:Electrophoresis, Polyacrylamide Gel; D004663:Encephalitis Virus, Eastern Equine; D005416:Flavivirus / Q000378:metabolism*; D015870:Gene Expression; D015151:Immunoblotting; D007668:Kidney; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D014162:Transfection; D014759:Viral Envelope Proteins / Q000096:biosynthesis* / Q000302:isolation & purification
chemical_list: D000911:Antibodies, Monoclonal; D046911:Macromolecular Substances; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C065696:glycoprotein E, Flavivirus