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Redirecting the substrate specificity of heparan sulfate 2-O-sulfotransferase by structurally guided mutagenesis.

Bethea HN, Xu D, Liu J, Pedersen LC
Proc Natl Acad Sci U S A 105 18724-9 (2008)
Cited: 35 times
EuropePMC logo PMID: 19022906

Abstract

Heparan sulfate (HS) is a polysaccharide involved in essential physiological functions from regulating cell growth to blood coagulation. HS biosynthesis involves multiple specialized sulfotransferases such as 2-O-sulfotransferase (2OST) that transfers the sulfo group to the 2-OH position of iduronic acid (IdoA) or glucuronic acid (GlcA) within HS. Here, we report the homotrimeric crystal structure of 2OST from chicken, in complex with 3'-phosphoadenosine 5'-phosphate. Structural based mutational analysis has identified amino acid residues that are responsible for substrate specificity. The mutant R189A only transferred sulfates to GlcA moieties within the polysaccharide whereas mutants Y94A and H106A preferentially transferred sulfates to IdoA units. Our results demonstrate the feasibility for manipulating the substrate specificity of 2OST to synthesize HS with unique sulfation patterns. This work will aid the development of an enzymatic approach to synthesize heparin-based therapeutics.

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  2. Crystal structures of MBP fusion proteins. Waugh DS. Protein Sci 25 559-571 (2016)
  3. Understanding the substrate specificity of the heparan sulfate sulfotransferases by an integrated biosynthetic and crystallographic approach. Liu J, Moon AF, Sheng J, Pedersen LC. Curr Opin Struct Biol 22 550-557 (2012)
  4. Sulfotyrosine residues: Interaction specificity determinants for extracellular protein-protein interactions. Stewart V, Ronald PC. J Biol Chem 298 102232 (2022)
  5. From Steroid and Drug Metabolism to Glycobiology, Using Sulfotransferase Structures to Understand and Tailor Function. Pedersen LC, Yi M, Pedersen LG, Kaminski AM. Drug Metab Dispos 50 1027-1041 (2022)

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Reviews citing this publication (6)

  1. Recent progress and applications in glycosaminoglycan and heparin research. Laremore TN, Zhang F, Dordick JS, Liu J, Linhardt RJ. Curr Opin Chem Biol 13 633-640 (2009)
  2. Recent advances in sulfotransferase enzyme activity assays. Paul P, Suwan J, Liu J, Dordick JS, Linhardt RJ. Anal Bioanal Chem 403 1491-1500 (2012)
  3. Use of biosynthetic enzymes in heparin and heparan sulfate synthesis. Chappell EP, Liu J. Bioorg Med Chem 21 4786-4792 (2013)
  4. Specific sides to multifaceted glycosaminoglycans are observed in embryonic development. Kramer KL. Semin Cell Dev Biol 21 631-637 (2010)
  5. The design and synthesis of new synthetic low-molecular-weight heparins. Chandarajoti K, Liu J, Pawlinski R. J Thromb Haemost 14 1135-1145 (2016)
  6. New tools for evaluating protein tyrosine sulfation and carbohydrate sulfation. Yeoh S, Bayliss R. Biochem J 475 3035-3037 (2018)

Articles citing this publication (19)



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