ArticleAnalytical and bioanalytical chemistry2014
Assays for determining heparan sulfate and heparin O-sulfotransferase activity and specificity.
Article in Analytical and bioanalytical chemistry, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 22 citations in OpenAlex.
- EstablishingBiodesign research · 2026Article
- Biosynthesis of animal-free recombinant chondroitin sulfate E using a functional chondroitin sulfotransferase in E. coli.Applied microbiology and biotechnology · 2024Article
- From Steroid and Drug Metabolism to Glycobiology, Using Sulfotransferase Structures to Understand and Tailor Function.Drug metabolism and disposition: the biological fate of chemicals · 2022Review
- Complete biosynthesis of a sulfated chondroitin in Escherichia coli.Nature communications · 2021Article
- Advances in the preparation and synthesis of heparin and related products.Drug discovery today · 2020Review
- Elucidating the unusual reaction kinetics of D-glucuronyl C5-epimerase.Glycobiology · 2020Article
- Heavy Heparin: A Stable Isotope-Enriched, Chemoenzymatically-Synthesized, Poly-Component Drug.Angewandte Chemie (International ed. in English) · 2019Article
- Engineered heparins as new anticoagulant drugs.Bioengineering & translational medicine · 2017Review
- Bioengineered heparins and heparan sulfates.Advanced drug delivery reviews · 2016Review
- Heparin and related polysaccharides: synthesis using recombinant enzymes and metabolic engineering.Applied microbiology and biotechnology · 2015Review
- Combinatorial one-pot chemoenzymatic synthesis of heparin.Carbohydrate polymers · 2015Article
- High cell density cultivation of recombinant Escherichia coli strains expressing 2-O-sulfotransferase and C5-epimerase for the production of bioengineered heparin.Applied biochemistry and biotechnology · 2015Article
Corrections and comments
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Authors and funding
7 authors at 2 institutions in 1 country.
Funding
Abstract
O-sulfotransferases (OSTs) are critical enzymes in the cellular biosynthesis of the biologically and pharmacologically important heparan sulfate and heparin. Recently, these enzymes have been cloned and expressed in bacteria for application in the chemoenzymatic synthesis of glycosaminoglycan-based drugs. OST activity assays have largely relied on the use of radioisotopic methods using [(35)S] 3'-phosphoadenosine-5'-phosphosulfate and scintillation counting. Herein, we examine alternative assays that are more compatible with a biomanufacturing environment. A high throughput microtiter-based approach is reported that relies on a coupled bienzymic colorimetric assay for heparan sulfate and heparin OSTs acting on polysaccharide substrates using arylsulfotransferase-IV and p-nitrophenylsulfate as a sacrificial sulfogroup donor. A second liquid chromatography-mass spectrometric assay, for heparan sulfate and heparin OSTs acting on structurally defined oligosaccharide substrates, is also reported that provides additional information on the number and positions of the transferred sulfo groups within the product. Together, these assays allow quantitative and mechanistic information to be obtained on OSTs that act on heparan sulfate and heparin precursors.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.