Tick-borne encephalitis virus envelope protein E-specific monoclonal antibodies for the study of low pH-induced conformational changes and immature virions

1995 ◽  
Vol 140 (2) ◽  
pp. 213-221 ◽  
Author(s):  
H. Holzmann ◽  
K. Stiasny ◽  
H. York ◽  
F. Dorner ◽  
C. Kunz ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Conformational Changes ◽  
Envelope Protein ◽  
Low Ph ◽  
Encephalitis Virus ◽  
Virus Envelope ◽  
Tick Borne Encephalitis ◽  
Virus Envelope Protein ◽  
Tick Borne Encephalitis Virus

scholarly journals N-linked glycan in tick-borne encephalitis virus envelope protein affects viral secretion in mammalian cells, but not in tick cells

2013 ◽  
Vol 94 (10) ◽  
pp. 2249-2258 ◽  
Author(s):  
Kentaro Yoshii ◽  
Natsumi Yanagihara ◽  
Mariko Ishizuka ◽  
Mizuki Sakai ◽  
Hiroaki Kariwa
Keyword(s):  
Envelope Protein ◽  
Mammalian Cells ◽  
Molecular Mechanisms ◽  
Consensus Sequence ◽  
Encephalitis Virus ◽  
Secretory Process ◽  
Virus Envelope ◽  
Tick Borne Encephalitis ◽  
Virus Envelope Protein ◽  
Tick Borne Encephalitis Virus

Tick-borne encephalitis virus (TBEV) is a zoonotic disease agent that causes severe encephalitis in humans. The envelope protein E of TBEV has one N-linked glycosylation consensus sequence, but little is known about the biological function of the N-linked glycan. In this study, the function of protein E glycosylation was investigated using recombinant TBEV with or without the protein E N-linked glycan. Virion infectivity was not affected after removing the N-linked glycans using N-glycosidase F. In mammalian cells, loss of glycosylation affected the conformation of protein E during secretion, reducing the infectivity of secreted virions. Mice subcutaneously infected with TBEV lacking protein E glycosylation showed no signs of disease, and viral multiplication in peripheral organs was reduced relative to that with the parental virus. In contrast, loss of glycosylation did not affect the secretory process of infectious virions in tick cells. Furthermore, inhibition of transport to the Golgi apparatus affected TBEV secretion in mammalian cells, but not in tick cells, indicating that TBEV was secreted through an unidentified pathway after synthesis in endoplasmic reticulum in tick cells. These results increase our understanding of the molecular mechanisms of TBEV maturation.

scholarly journals Mapping of Functional Elements in the Stem-Anchor Region of Tick-Borne Encephalitis Virus Envelope Protein E

1999 ◽  
Vol 73 (7) ◽  
pp. 5605-5612 ◽  
Author(s):  
Steven L. Allison ◽  
Karin Stiasny ◽  
Konrad Stadler ◽  
Christian W. Mandl ◽  
Franz X. Heinz
Keyword(s):  
Amino Acids ◽  
Membrane Fusion ◽  
Envelope Protein ◽  
Molecular Mechanisms ◽  
Low Ph ◽  
Encephalitis Virus ◽  
E Proteins ◽  
Functional Elements ◽  
Tick Borne Encephalitis ◽  
Tick Borne Encephalitis Virus

ABSTRACT Envelope protein E of the flavivirus tick-borne encephalitis virus mediates membrane fusion, and the structure of the N-terminal 80% of this 496-amino-acid-long protein has been shown to differ significantly from that of other viral fusion proteins. The structure of the carboxy-terminal 20%, the stem-anchor region, is not known. It contains sequences that are important for membrane anchoring, interactions with prM (the precursor of membrane protein M) during virion assembly, and low-pH-induced structural changes associated with the fusion process. To identify specific functional elements in this region, a series of C-terminal deletion mutants were constructed and the properties of the resulting truncated recombinant E proteins were examined. Full-length E proteins and proteins lacking the second of two predicted transmembrane segments were secreted in a particulate form when coexpressed with prM, whereas deletion of both segments resulted in the secretion of soluble homodimeric E proteins. Sites located within a predicted α-helical region of the stem (amino acids 431 to 449) and the first membrane-spanning region (amino acids 450 to 472) were found to be important for the stability of the prM-E heterodimer but not essential for prM-mediated intracellular transport and secretion of soluble E proteins. A separate site in the stem, also corresponding to a predicted α-helix (amino acids 401 to 413), was essential for the conversion of soluble protein E dimers to a homotrimeric form upon low-pH treatment, a process resembling the transition to the fusogenic state in whole virions. This functional mapping will aid in the understanding of the molecular mechanisms of membrane fusion and virus assembly.

scholarly journals Membrane Interactions of the Tick-Borne Encephalitis Virus Fusion Protein E at Low pH

2002 ◽  
Vol 76 (8) ◽  
pp. 3784-3790 ◽  
Author(s):  
Karin Stiasny ◽  
Steven L. Allison ◽  
Juliane Schalich ◽  
Franz X. Heinz
Keyword(s):  
Fusion Protein ◽  
Membrane Binding ◽  
Low Ph ◽  
Encephalitis Virus ◽  
Soluble Form ◽  
Viral Fusion ◽  
Sequence Element ◽  
Viral Fusion Protein ◽  
Tick Borne Encephalitis ◽  
Tick Borne Encephalitis Virus

ABSTRACT Membrane fusion of the flavivirus tick-borne encephalitis virus is triggered by the mildly acidic pH of the endosome and is mediated by envelope protein E, a class II viral fusion protein. The low-pH trigger induces an oligomeric rearrangement in which the subunits of the native E homodimers dissociate and the monomeric subunits then reassociate into homotrimers. Here we provide evidence that membrane binding is mediated by the intermediate monomeric form of E, generated by low-pH-induced dissociation of the dimer. Liposome coflotation experiments revealed that association with target membranes occurred only when liposomes were present at the time of acidification, whereas pretreating virions at low pH in the absence of membranes resulted in the loss of their ability to stably attach to liposomes. With the cleavable cross-linker ethylene glycolbis(succinimidylsuccinate), it was shown that a truncated soluble form of the E protein (sE) could bind to membranes only when the dimers were free to dissociate at low pH, and binding could be blocked by a monoclonal antibody that recognizes the fusion peptide, which is at the distal tip of the E monomer but is buried in the native dimer. Surprisingly, analysis of the membrane-associated sE proteins revealed that they had formed trimers. This was unexpected because this protein lacks a sequence element in the C-terminal stem-anchor region, which was shown to be essential for trimerization in the absence of a target membrane. It can therefore be concluded that the formation of a trimeric form of sE is facilitated by membrane binding. Its stability is apparently maintained by contacts between the ectodomains only and is not dependent on sequence elements in the stem-anchor region as previously assumed.  

scholarly journals Characterization of Monoclonal Antibodies against Hokkaido Strain Tick-Borne Encephalitis Virus

2000 ◽  
Vol 44 (6) ◽  
pp. 533-536 ◽  
Author(s):  
Kimiyo Komoro ◽  
Daisuke Hayasaka ◽  
Tetsuya Mizutani ◽  
Hiroaki Kariwa ◽  
Ikuo Takashima
Keyword(s):  
Monoclonal Antibodies ◽  
Encephalitis Virus ◽  
Tick Borne Encephalitis ◽  
Tick Borne Encephalitis Virus
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