title: Identification of the cell binding domain in Nipah virus G glycoprotein using a phage display system.
abstract: Nipah virus (NiV) is a highly pathogenic zoonotic paramyxovirus with unusual broad host tropism and is designated as a Category C pathogen by the U.S. National Institute of Allergy and Infectious Diseases. NiV infection is initiated after binding of the viral G glycoprotein to the host cell receptor. The aim of this study was to map the NiV G glycoprotein cell binding domain using a phage display system. The NiV G extracellular domain was truncated and displayed as attachment proteins on M13 phage g3p minor coat protein. The binding efficiency of recombinant phages displaying different regions of NiV G to mammalian cells was evaluated. Results showed that regions of NiV G consisting of amino acids 396-602 (recombinant phage G4) and 498-602 (recombinant phage G5) demonstrated the highest binding to both Vero (5.5×103cfu/ml and 5.6×103cfu/ml) and THP-1 cells (3.5×103cfu/ml and 2.9×103cfu/ml). However, the binding of both of these recombinant phages to THP-1 cells was significantly lower than to Vero cells, and this could be due to the lack of primary host cell receptor expression on THP-1 cells. Furthermore, the binding between these two recombinant phages was competitive suggesting that there was a common host cell attachment site. This study employed an approach that is suitable for use in a biosafety level 2 containment laboratory without the need to use live virus to show that NiV G amino acids 498-602 play an important role for attachment to host cells.
authors.affiliation: Department of Medical Microbiology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia.
authors.forename: Chui-Wan
authors.initials: CW
authors.lastname: Lam
authors.affiliation: Department of Medical Microbiology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia; Tropical Infectious Diseases Research and Education Centre (TIDREC), University of Malaya, 50603 Kuala Lumpur, Malaysia.
authors.forename: Sazaly
authors.initials: S
authors.lastname: AbuBakar
authors.affiliation: Department of Medical Microbiology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia; Tropical Infectious Diseases Research and Education Centre (TIDREC), University of Malaya, 50603 Kuala Lumpur, Malaysia. Electronic address: changliyen@um.edu.my.
authors.forename: Li-Yen
authors.initials: LY
authors.lastname: Chang
mesh_terms: D000818:Animals; D017104:Bacteriophage M13 / Q000235:genetics; D001665:Binding Sites; D002460:Cell Line; D002522:Chlorocebus aethiops; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D045405:Nipah Virus / Q000502:physiology*; D019151:Peptide Library; D011485:Protein Binding; D000072417:Protein Domains; D015678:Viral Structural Proteins / Q000378:metabolism*; D053585:Virus Attachment
chemical_list: D006023:Glycoproteins; D019151:Peptide Library; D015678:Viral Structural Proteins
keywords: Glycoprotein; Host cell receptor; Infectious disease; Nipah virus; Phage display