Dengue virus type 2 envelope protein displayed as recombinant phage attachment protein reveals potential cell binding sites

Abd-Jamil, J.; Cheah, C.Y.; AbuBakar, S. (2008) Dengue virus type 2 envelope protein displayed as recombinant phage attachment protein reveals potential cell binding sites. Protein Engineering Design & Selection, 21 (10). pp. 605-611. ISSN 1741-0134

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    A method to map the specific site on dengue virus envelope protein (E) that interacts with cells and a neutralizing antibody is developed using serially truncated dengue virus type 2 (DENV-2) E displayed on M13 phages as recombinant E-g3p fusion proteins. Recombinant phages displaying the truncated E consisting of amino acids 297-423 (EB2) and amino acids 379-423 (EB4) were neutralized by DENV-2 patient sera and the DENV-2 E-specific 3H5-1 monoclonal antibodies suggesting that the phages retained the DENV-2 E antigenic properties. The EB4 followed by EB2 recombinant phages bound the most to human monocytes (THP-1), African green monkey kidney (Vero) cells, mosquito (C6/36) cells, ScFv specific against E and C6/36 cell proteins. Two potential cell attachment sites were mapped to loop I (amino acids 297 to 312) and loop II (amino acids 379-385) of the DENV-2 E using the phage-displayed truncated DENV-2 E fragments and by the analysis of the E structure. Loop II was present only in EB4 recombinant phages. There was no competition for binding to C6/36 cell proteins between EB2 and EB4 phages. Loop I and loop II are similar to the sub-complex specific and type-specific neutralizing monoclonal antibody binding sites, respectively. Hence, it is proposed that binding and entry of DENV involves the interaction of loop I to cell surface glycosaminoglycan-motif and a subsequent highly specific interaction involving loop II with other cell proteins. The phage displayed truncated DENV-2 E is a powerful and useful method for the direct determination of DENV-2 E cell binding sites.

    Item Type: Article
    1. Abd-Jamil, J.
    2. Cheah, C.Y.
    3. AbuBakar, S.(Department of Medical Microbiology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, MALAYSIA)
    Journal or Publication Title: Protein Engineering Design & Selection
    Additional Information: Abd-Jamil, Juraina Cheah, Chen-Yee AbuBakar, Sazaly eng Research Support, Non-U.S. Gov't England 2008/08/02 09:00 Protein Eng Des Sel. 2008 Oct;21(10):605-11. Epub 2008 Jul 30.
    Uncontrolled Keywords: Animals Antibody Specificity Bacteriophage M13/immunology/metabolism/*physiology Binding Sites Binding, Competitive Cell Line Cells/immunology/*metabolism Culicidae/immunology/metabolism Dengue Virus/immunology/metabolism/*physiology Gene Deletion Gene Expression Regulation Humans Immunoglobulin Variable Region/immunology Neutralization Tests Protein Structure, Tertiary Recombinant Fusion Proteins/chemistry/genetics/immunology/*metabolism Viral Envelope Proteins/chemistry/genetics/immunology/*metabolism *Virus Attachment
    Subjects: R Medicine
    Divisions: Faculty of Medicine
    Depositing User: Mr Jenal S
    Date Deposited: 08 Nov 2012 12:04
    Last Modified: 07 Nov 2017 16:30

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