Novel anti-dengue monoclonal antibody recognizing conformational structure of the prM-E heterodimeric complex of dengue virus.
An interaction between the premembrane (prM) and envelope (E) glycoproteins as prM-E heterodimer is required for proper folding and transport of E during the formation and release of new flaviviral progeny. More evidence, however, is needed to confirm this interaction of prM and E during dengue virus replication. In this study, 2E11, a mouse monoclonal antibody (Mab) that specifically recognizes dengue prM-E heterodimeric complex in either intracellular or secreted dengue virions, was generated and characterized. In immunofluorescence and immuno-pull down assays, the Mab 2E11 recognized an epitope present in 293T transfectants that co-expressed prM and the full-length form of E in cis and in trans, but it failed to react with prM or E protein expressed individually. The reactivity of Mab 2E11 was diminished in transfected cells that co-express prM together with a truncated form of E lacking the 84-residue stretch at the C-terminal transmembrane region, presumably essential for prM and E interaction. The Mab 2E11 described in this study is a novel Mab with a unique capability in detecting the conformational structure of prM-E heterodimeric complex of dengue virus. It will be a new biological tool for identification and characterization of dengue prM-E heterodimer as well as virus maturation and export.
Medical Biotechnology Unit, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, Bangkok, Thailand.
Chunya
C
Puttikhunt
Poonsook
P
Keelapang
Nuanpan
N
Khemnu
Nopporn
N
Sittisombut
Watchara
W
Kasinrerk
Prida
P
Malasit
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D002460:Cell Line; D003715:Dengue / Q000276:immunology / Q000517:prevention & control / Q000821:virology; D003716:Dengue Virus / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D019281:Dimerization; D006023:Glycoproteins; D051379:Mice; D008807:Mice, Inbred BALB C; D011487:Protein Conformation; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000276:immunology*
D000911:Antibodies, Monoclonal; C051821:E-glycoprotein, Dengue virus type 2; D006023:Glycoproteins; D014759:Viral Envelope Proteins; C074932:prM protein, Flavivirus