Selection of rotavirus VP4 cell receptor binding domains for MA104 cells using a phage display library.
Rotavirus infection of host cells, like other viruses, is a complex process that has not been fully elucidated, and much attention has been focused on the regions of the viral attachment protein, VP4, that are involved in binding to the cellular receptor. In this study, phage display technology was employed to generate a g3p VP4 gene-targeted phage display peptide library using the porcine rotavirus strain CRW8, and a method was optimised for panning this library on adherent MA104 cells to identify receptor binding domains. Recombinant phage that displayed expressed peptides from both the rotavirus VP4 trypsin cleavage products VP8* and VP5* were selected, and while some of the phage clones contained insert sequences from regions of VP4 implicated previously in cell binding and infection, new domains were also identified. In all, four regions within VP8* and six regions of VP5* were selected by panning. To our knowledge, this paper is the first description of using a gene-targeted phage display library to identify receptor binding domains on viral proteins.
Jolly
C L
CL
Department of Microbiology and Immunology, University of Melbourne, Parkville Victoria 3010, Melbourne, Australia. rota@microbiology.unimelb.edu.au
Huang
J A
JA
Holmes
I H
IH
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002213:Capsid / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D036022:Capsid Proteins; D002460:Cell Line; D002522:Chlorocebus aethiops; D003090:Coliphages / Q000235:genetics; D008969:Molecular Sequence Data; D019151:Peptide Library; D011991:Receptors, Virus / Q000737:chemistry / Q000378:metabolism*; D012401:Rotavirus / Q000235:genetics / Q000378:metabolism*; D013552:Swine; D014357:Trypsin
D036022:Capsid Proteins; D019151:Peptide Library; D011991:Receptors, Virus; C060339:VP4 protein, Rotavirus; D014357:Trypsin