Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
SHR Neuro Krebs Kardio Lipid Stoffw Microb
Wille, I; Rek, A; Krenn, E; Kungl, AJ.
Biophysical investigation of human heparan sulfate D-glucosaminyl 3-O-sulfotransferase-3A: a mutual effect of enzyme oligomerisation and glycosaminoglycan ligand binding.
Biochim Biophys Acta. 2007; 1774(11): 1470-6.
Doi: 10.1016/j.bbapap.2007.08.023
Web of Science
PubMed
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- Co-Autor*innen der Med Uni Graz
- Jantscher-Krenn Evelyn
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- Abstract:
- 3-O-sulfation of heparan sulfate (HS) is the rarest modification within heparan sulfate biosynthesis resulting in unique biological activities. Heparan sulfate d-glucosaminyl 3-O-sulfotransferase-3A (3-OST-3A) (EC 2.8.2.23) generates a binding site for the envelope glycoprotein D (gD) of herpes simplex virus 1. We have expressed the sulfotransferase domain of the human heparan sulfate 3-OST-3A isoform in Escherichia coli and subsequently purified the active enzyme which was found to be present as an oligomer under nonreducing conditions. The activity of the enzyme was tested by a novel gD-dependent gel mobility assay. A biophysical characterisation of 3-OST-3A was performed to study ligand binding and ligand-induced structural changes. Interestingly, the natural substrate HS did not cause a secondary structural change in the enzyme, whereas heparin and chondroitin sulfate did, both of which also exhibited similar high affinity binding to 3-OST-3A compared to HS as detected by isothermal fluorescence titrations. In cross-link assays, only HS was found to induce high molecular aggregates of 3-OST-3A whereas other GAG ligands did not or even inhibited enzyme oligomerisation like the K5 polysaccharide, which was nevertheless found to bind to the enzyme. We therefore conclude that since 3-OST-3A is able to bind also non-substrate GAG ligands with high affinity, discrimination among ligands is triggered by protein oligomerisation.
- Find related publications in this database (using NLM MeSH Indexing)
- Circular Dichroism - administration & dosage
- Escherichia coli - genetics
- Glycosaminoglycans - metabolism
- Humans - administration & dosage
- Isoenzymes - chemistry, genetics, metabolism
- Ligands - administration & dosage
- Protein Binding - administration & dosage
- Protein Structure, Tertiary - administration & dosage
- Recombinant Proteins - chemistry, genetics, metabolism
- Sulfotransferases - chemistry, genetics, metabolism
- Find related publications in this database (Keywords)
- fluorescence
- circular dichroism
- folding
- enzyme
- biosynthesis