Humans Express Seven Heparan Sulfate (HS) 3-O-Sulfotransferases that Differ in Substrate Specificity and Tissue Expression. Although Genetic Studies Have Indicated that 3-O-Sulfated HS Modulates Many Biological Processes, Ligand Requirements for Proteins Engaging with HS Modified by 3-O-Sulfate (3-OS) Have Been Difficult to Determine. in Particular, the Context in Which the 3-OS Group Needs to Be Presented for Binding is Largely Unknown. We Describe Herein a Modular Synthetic Approach that Can Provide Structurally Diverse HS Oligosaccharides with and Without 3-OS. the Methodology Was Employed to Prepare 27 Hexasaccharides that Were Printed as a Glycan Microarray to Examine Ligand Requirements of a Wide Range of HS-Binding Proteins. the Binding Selectivity of Antithrombin-III (AT-III) Compared Well with Anti-Factor Xa Activity Supporting Robustness of the Array Technology. Many of the Other Examined HS-Binding Proteins Required an IdoA2SGlcNS3S6S Sequon for Binding But Exhibited Variable Dependence for the 2-OS and 6-OS Moieties, and a GlcA or IdoA2S Residue Neighboring the Central GlcNS3S. the HS Oligosaccharides Were Also Examined as Inhibitors of Cell Entry by Herpes Simplex Virus Type 1, Which, Surprisingly, Showed a Lack of Dependence of 3-OS, Indicating that, Instead of Glycoprotein D (GD), They Competitively Bind to GB and GC. the Compounds Were Also Used to Examine Substrate Specificities of Heparin Lyases, Which Are Enzymes Used for Depolymerization of HS/heparin for Sequence Determination and Production of Therapeutic Heparins. It Was Found that Cleavage by Lyase II is Influenced by 3-OS, While Digestion by Lyase I is Only Affected by 2-OS. Lyase III Exhibited Sensitivity to Both 3-OS and 2-OS.
P. Chopra et al., "The 3-O-Sulfation of Heparan Sulfate Modulates Protein Binding and Lyase Degradation," Proceedings of the National Academy of Sciences of the United States of America, vol. 118, no. 3, article no. e2012935118, National Academy of Sciences, Jan 2021.
The definitive version is available at https://doi.org/10.1073/PNAS.2012935118
Chemical and Biochemical Engineering
Keywords and Phrases
3-O-sulfation; Anti-Factor Xa; Glycan microarray; Heparin lyases; Herpes simplex virus 1
International Standard Serial Number (ISSN)
Article - Journal
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13 Jan 2021
National Institutes of Health, Grant R01HL151617