| Title | Characterization of a heparan sulfate octasaccharide that binds to herpes simplex virus type 1 glycoprotein D. | | Author(s) | Liu J, Shriver Z, Pope RM, Thorp SC, Duncan MB, Copeland RJ, Raska CS, Yoshida K, Eisenberg RJ, Cohen G, Linhardt RJ, Sasisekharan R | | Institution | Division of Medicinal Chemistry and Natural Products, School of Pharmacy, University of North Carolina, Chapel Hill, North Carolina 27599, USA. jian_liu@unc.edu | | Source | J Biol Chem 2002 Sep 6; 277(36):33456-67. | | MeSH | Animals
Chromatography, High Pressure Liquid
Disaccharides
Electrophoresis, Capillary
Insects
Kinetics
Models, Chemical
Polysaccharides
Protein Binding
Protein Isoforms
Protein Structure, Tertiary
Recombinant Fusion Proteins
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.
Spectrometry, Mass, Electrospray Ionization
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Time Factors
Ultraviolet Rays
Viral Envelope Proteins
| | Abstract | Herpes simplex virus type 1 utilizes cell surface heparan sulfate as receptors to infect target cells. The unique heparan sulfate saccharide sequence offers the binding site for viral envelope proteins and plays critical roles in assisting viral infections. A specific 3-O-sulfated heparan sulfate is known to facilitate the entry of herpes simplex virus 1 into cells. The 3-O-sulfated heparan sulfate is generated by the heparan sulfate d-glucosaminyl-3-O-sulfotransferase isoform 3 (3-OST-3), and it provides binding sites for viral glycoprotein D (gD). Here, we report the purification and structural characterization of an oligosaccharide that binds to gD. The isolated gD-binding site is an octasaccharide, and has a binding affinity to gD around 18 microm, as determined by affinity coelectrophoresis. The octasaccharide was prepared and purified from a heparan sulfate oligosaccharide library that was modified by purified 3-OST-3 enzyme. The molecular mass of the isolated octasaccharide was determined using both nanoelectrospray ionization mass spectrometry and matrix-assisted laser desorption/ionization mass spectrometry. The results from the sequence analysis suggest that the structure of the octasaccharide is a heptasulfated octasaccharide. The proposed structure of the octasaccharide is DeltaUA-GlcNS-IdoUA2S-GlcNAc-UA2S-GlcNS-IdoUA2S-GlcNH(2)3S6S. Given that the binding of 3-O-sulfated heparan sulfate to gD can mediate viral entry, our results provide structural information about heparan sulfate-assisted viral entry. | | Language | eng | | Pub Type(s) | Journal Article
| | PubMed ID | 12080045 |
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