3F5F

Crystal structure of heparan sulfate 2-O-sulfotransferase from gallus gallus as a maltose binding protein fusion.

Summary for 3F5F

• Entry DOI: 10.2210/pdb3f5f/pdb
• Related PRD ID: PRD_900001
• Descriptor: Maltose-binding periplasmic protein, Heparan sulfate 2-O-sulfotransferase 1, alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose, ADENOSINE-3'-5'-DIPHOSPHATE, ... (4 entities in total)
• Functional Keywords: maltose binding protein, fusion, heparan sulfate biosynthesis, sulfotransferase, glycoprotein, golgi apparatus, membrane, signal-anchor, transferase, transmembrane, periplasm, sugar transport, transport
• Biological source: Escherichia coli K-12 (bantam,chickens); More
• Cellular location: Golgi apparatus membrane ; Single-pass type II membrane protein : Q76KB1
• Total number of polymer chains: 1
• Total formula weight: 75055.83

Authors

Bethea, H.N.,Xu, D.,Liu, J.,Pedersen, L.C. (deposition date: 2008-11-03, release date: 2008-12-16, Last modification date: 2024-10-16)

Primary citation

Bethea, H.N.,Xu, D.,Liu, J.,Pedersen, L.C.
Redirecting the substrate specificity of heparan sulfate 2-O-sulfotransferase by structurally guided mutagenesis.
Proc.Natl.Acad.Sci.USA, 105:18724-18729, 2008

PubMed 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.

PubMed: 19022906
DOI: 10.1073/pnas.0806975105

Experimental method

X-RAY DIFFRACTION (2.65 Å)