Multiple heparin binding domains of respiratory syncytial virus G mediate binding to mammalian cells

• Published in: Archives of virology Vol. 148; no. 10; pp. 1987 - 2003
• Main Authors: SHIELDS, B, MILLS, J, GHILDYAL, R, GOOLEY, P, MEANGER, J
• Format: Journal Article
• Language: English
• Published: Wien Springer 01.10.2003; New York, NY Springer Nature B.V
• Subjects: Amino Acid Sequence; Amino acids; Animals; Biological and medical sciences; CHO Cells; Cricetinae; Dengue fever; Encephalitis; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Viral; Heparan sulfate; Heparin - metabolism; Herpes viruses; Human viral diseases; Humans; Infectious diseases; Medical sciences; Microbiology; Miscellaneous; Molecular Sequence Data; Mutation; Proteins; Recombinant Proteins - metabolism; Respiratory syncytial virus; Respiratory Syncytial Viruses - metabolism; Respiratory Syncytial Viruses - pathogenicity; Sequence Alignment; Tumor Cells, Cultured; Viral diseases; Viral infections; Viral Plaque Assay; Viral Proteins - chemistry; Viral Proteins - genetics; Viral Proteins - metabolism; Virology; Human respiratory syncytial virus; Multiple; Pneumovirinae; Respiratory system; Paramyxoviridae; Virus; Respiratory tract; Mononegavirales; Vertebrata; Mammalia; Glycosaminoglycan; Heparin; Pneumovirus
• ISSN: 0304-8608; 1432-8798
• DOI: 10.1007/s00705-003-0139-0

Respiratory syncytial virus (RSV) G glycoprotein mediates cell attachment through surface glycosaminoglycans (GAGs). Feldman et al. [10] suggested that specific basic amino acids in residues 184-198 of G defined a critical heparin binding domain (HBD). To further define the G HBD we made a series of truncated G proteins expressed in Escherichia coli. G88 (G residues 143-231), bound to HEp-2 cells in a dose dependent manner and binding was inhibited >99% with heparin. Cell binding of G88 was unaltered by alanine substitution mutagenesis of all basic amino acids in Feldman's region 184-198. A G88 variant truncated beyond residue 198, G58, and G58 fully alanine substituted in the region 184-198, G58A6, bound to HEp-2 cells about half as well and 100-fold less well than G88, respectively. G88 and all alanine substitution mutants of G88 inhibited RSV plaque formation by 50% (ID(50)) at concentrations of approximately 50 nM; the ID(50) of G58 was approximately 425 nM while G58A6 had an ID(50) >1600 nM. These data show that the G HBD includes as much as residues 187-231, that there is redundancy beyond the previously described HBD, and that the cell-binding and virus infectivity-blocking functions of these recombinant G proteins were closely linked and required at least one HBD.