The Library
Structural analysis of the adenovirus type 2 E3/19K protein using mutagenesis and a panel of conformation-sensitive monoclonal antibodies
Tools
Menz, Beatrice, Sester, Martina, Koebernick, Katja, Schmid, Ralf and Burgert, Hans-Gerhard (2008) Structural analysis of the adenovirus type 2 E3/19K protein using mutagenesis and a panel of conformation-sensitive monoclonal antibodies. Molecular Immunology, Vol.46 (No.1). pp. 16-26. doi:10.1016/j.molimm.2008.06.019 ISSN 0161-5890.
|
PDF
WRAP_Burgert_Paper__Menz__revised__Figs10608.pdf - Requires a PDF viewer. Download (2347Kb) |
Official URL: http://dx.doi.org/10.1016/j.molimm.2008.06.019
Abstract
The E3/19K protein of human adenovirus type 2 (Ad2) was the first viral protein shown to interfere with antigen presentation. This 25 kDa transmembrane glycoprotein binds to major histocompatibility complex (MHC) class I molecules in the endoplasmic reticulum (ER), thereby preventing transport of newly synthesized peptide–MHC complexes to the cell surface and consequently T cell recognition. Recent data suggest that E3/19K also sequesters MHC class I like ligands intracellularly to suppress natural killer (NK) cell recognition. While the mechanism of ER retention is well understood, the structure of E3/19K remains elusive. To further dissect the structural and antigenic topography of E3/19K we carried out site-directed mutagenesis and raised monoclonal antibodies (mAbs) against a recombinant version of Ad2 E3/19K comprising the lumenal domain followed by a C-terminal histidine tag. Using peptide scanning, the epitopes of three mAbs were mapped to different regions of the lumenal domain, comprising amino acids 3–13, 15–21 and 41–45, respectively. Interestingly, mAb 3F4 reacted only weakly with wild-type E3/19K, but showed drastically increased binding to mutant E3/19K molecules, e.g. those with disrupted disulfide bonds, suggesting that 3F4 can sense unfolding of the protein. MAb 10A2 binds to an epitope apparently buried within E3/19K while that of 3A9 is exposed. Secondary structure prediction suggests that the lumenal domain contains six β-strands and an α-helix adjacent to the transmembrane domain. Interestingly, all mAbs bind to non-structured loops. Using a large panel of E3/19K mutants the structural alterations of the mutations were determined. With this knowledge the panel of mAbs will be valuable tools to further dissect structure/function relationships of E3/19K regarding down regulation of MHC class I and MHC class I like molecules and its effect on both T cell and NK cell recognition.
Item Type: | Journal Article | ||||
---|---|---|---|---|---|
Subjects: | Q Science > QR Microbiology > QR355 Virology | ||||
Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) > Biological Sciences ( -2010) | ||||
Library of Congress Subject Headings (LCSH): | Adenoviruses, Immunology, Antigens | ||||
Journal or Publication Title: | Molecular Immunology | ||||
Publisher: | Elsevier | ||||
ISSN: | 0161-5890 | ||||
Official Date: | November 2008 | ||||
Dates: |
|
||||
Volume: | Vol.46 | ||||
Number: | No.1 | ||||
Page Range: | pp. 16-26 | ||||
DOI: | 10.1016/j.molimm.2008.06.019 | ||||
Status: | Peer Reviewed | ||||
Access rights to Published version: | Open Access (Creative Commons) |
Request changes or add full text files to a record
Repository staff actions (login required)
View Item |
Downloads
Downloads per month over past year