3UAN

Crystal structure of 3-O-sulfotransferase (3-OST-1) with bound PAP and heptasaccharide substrate

Summary for 3UAN

• Entry DOI: 10.2210/pdb3uan/pdb
• Related: 1T8T 1T8U 1ZRH
• Descriptor: Heparan sulfate glucosamine 3-O-sulfotransferase 1, 2-acetamido-2-deoxy-6-O-sulfo-alpha-D-glucopyranose-(1-4)-beta-D-glucopyranuronic acid-(1-4)-2-deoxy-6-O-sulfo-2-(sulfoamino)-alpha-D-glucopyranose-(1-4)-2-O-sulfo-alpha-L-idopyranuronic acid-(1-4)-2-deoxy-6-O-sulfo-2-(sulfoamino)-alpha-D-glucopyranose-(1-4)-beta-D-glucopyranuronic acid, 2-acetamido-2-deoxy-6-O-sulfo-alpha-D-glucopyranose-(1-4)-beta-D-glucopyranuronic acid-(1-4)-2-deoxy-6-O-sulfo-2-(sulfoamino)-alpha-D-glucopyranose-(1-4)-2-O-sulfo-alpha-L-idopyranuronic acid-(1-4)-2-deoxy-6-O-sulfo-2-(sulfoamino)-alpha-D-glucopyranose, ... (6 entities in total)
• Functional Keywords: sulfotransferase, heparan sulfate, alpha/beta motif, co-factor paps/pap, heparan sulfate oligosaccharides, golgi-localized, transferase
• Biological source: Mus musculus (mouse)
• Cellular location: Golgi apparatus lumen (Probable): O35310
• Total number of polymer chains: 2
• Total formula weight: 66784.72

Authors

Moon, A.F.,Xu, Y.,Woody, S.M.,Krahn, J.M.,Linhardt, R.J.,Liu, J.,Pedersen, L.C. (deposition date: 2011-10-21, release date: 2012-04-04, Last modification date: 2024-11-20)

Primary citation

Moon, A.F.,Xu, Y.,Woody, S.M.,Krahn, J.M.,Linhardt, R.J.,Liu, J.,Pedersen, L.C.
Dissecting the substrate recognition of 3-O-sulfotransferase for the biosynthesis of anticoagulant heparin.
Proc.Natl.Acad.Sci.USA, 109:5265-5270, 2012

PubMed Abstract: Heparin is a polysaccharide-based natural product that is used clinically as an anticoagulant drug. Heparan sulfate 3-O-sulfotransferase (3-OST) is an enzyme that transfers a sulfo group to the 3-OH position of a glucosamine unit. 3-OST is present in multiple isoforms, and the polysaccharides modified by these different isoforms perform distinct biological functions. 3-OST isoform 1 (3-OST-1) is the key enzyme for the biosynthesis of anticoagulant heparin. Here, we report the crystal structure of the ternary complex of 3-OST-1, 3'-phosphoadenosine 5'-phosphate, and a heptasaccharide substrate. Comparisons to previously determined structures of 3-OST-3 reveal unique binding modes used by the different isoforms of 3-OST for distinguishing the fine structures of saccharide substrates. Our data demonstrate that the saccharide substrates display distinct conformations when interacting with the different 3-OST isoforms. Site-directed mutagenesis data suggest that several key amino residues, including Lys259, Thr256, and Trp283 in 3-OST-3 and Arg268 in 3-OST-1, play important roles in substrate binding and specificity between isoforms. These results deepen our understanding of the biosynthetic mechanism of heparan sulfate and provide structural information for engineering enzymes for an enhanced biosynthetic approach to heparin production.

PubMed: 22431632
DOI: 10.1073/pnas.1117923109

Experimental method

X-RAY DIFFRACTION (1.844 Å)