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Journal of Virology, August 1999, p. 6610-6617, Vol. 73, No. 8
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Identification of a Linear Heparin Binding Domain for Human Respiratory Syncytial Virus Attachment Glycoprotein G

Steven A. Feldman,1,* R. Michael Hendry,2 and Judy A. Beeler1

Laboratory of Pediatric and Respiratory Virus Diseases, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland,1 and Viral and Rickettsial Diseases Laboratory, California Department of Health Services, Berkeley, California2

Received 19 October 1998/Accepted 10 April 1999

Respiratory syncytial virus (RSV) is the leading cause of lower respiratory tract disease in infants and young children worldwide. Infection is mediated, in part, by an initial interaction between attachment protein (G) and a highly sulfated heparin-like glycosaminoglycan (Gag) located on the cell surface. Synthetic overlapping peptides derived from consensus sequences of the G protein ectodomain from both RSV subgroups A and B were tested by heparin-agarose affinity chromatography for their abilities to bind heparin. This evaluation identified a single linear heparin binding domain (HBD) for RSV subgroup A (184Aright-arrowT198) and B (183Kright-arrowK197). The binding of these peptides to Vero cells was inhibited by heparin. Peptide binding to two CHO cell mutants (pgsD-677 and pgsA-745) deficient in heparan sulfate or total Gag synthesis was decreased 50% versus the parental cell line, CHO-K1, and decreased an average of 87% in the presence of heparin. The RSV-G HBD peptides were also able to inhibit homologous and heterologous virus infectivity of Vero cells. These results indicate that the sequence 184A/183Kright-arrow198T/K197 for RSV subgroups A and B, respectively, defines an important determinant of RSV-G interactions with heparin.

* Corresponding author. Mailing address: Food and Drug Administration, Center for Biologics Evaluation and Research, Building 29A, 3B-05, HFM 463, 1401 Rockville Pike, Rockville, MD 20852-1448. Phone: (301) 827-1939. Fax: (301) 496-1810. E-mail: feldmans{at}cber.fda.gov.

Journal of Virology, August 1999, p. 6610-6617, Vol. 73, No. 8
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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