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

B Shields, J Mills, R Ghildyal, P Gooley, J Meanger

Research output: Contribution to journalArticle

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Abstract

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.

Original languageEnglish
Pages (from-to)1987-2003
Number of pages17
JournalArchives of Virology
Volume148
Issue number10
DOIs
Publication statusPublished - 2003
Externally publishedYes

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Respiratory Syncytial Viruses
Heparin
Alanine
Basic Amino Acids
GTP-Binding Proteins
Virus Attachment
Glycosaminoglycans
Recombinant Proteins
Mutagenesis
Escherichia coli

Cite this

Shields, B ; Mills, J ; Ghildyal, R ; Gooley, P ; Meanger, J. / Multiple heparin binding domains of respiratory syncytial virus G mediate binding to mammalian cells. In: Archives of Virology. 2003 ; Vol. 148, No. 10. pp. 1987-2003.
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Multiple heparin binding domains of respiratory syncytial virus G mediate binding to mammalian cells. / Shields, B; Mills, J; Ghildyal, R; Gooley, P; Meanger, J.

In: Archives of Virology, Vol. 148, No. 10, 2003, p. 1987-2003.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Multiple heparin binding domains of respiratory syncytial virus G mediate binding to mammalian cells

AU - Shields, B

AU - Mills, J

AU - Ghildyal, R

AU - Gooley, P

AU - Meanger, J

PY - 2003

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AB - 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.

KW - Amino Acid Sequence

KW - Animals

KW - CHO Cells

KW - Cricetinae

KW - Gene Expression Regulation, Viral

KW - Heparin

KW - Humans

KW - Molecular Sequence Data

KW - Mutation

KW - Recombinant Proteins

KW - Respiratory Syncytial Viruses

KW - Sequence Alignment

KW - Tumor Cells, Cultured

KW - Viral Plaque Assay

KW - Viral Proteins

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

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DO - 10.1007/s00705-003-0139-0

M3 - Article

VL - 148

SP - 1987

EP - 2003

JO - Archiv fur die gesamte Virusforschung

JF - Archiv fur die gesamte Virusforschung

SN - 0304-8608

IS - 10

ER -