A role of Lys614 in the sulfotransferase activity of human heparan sulfate N‐deacetylase/N‐sulfotransferase

FEBS Letters

Kakuta, Yoshimitsu²  Wall, Frances E.¹  Pedersen, Lee G.¹  Sueyoshi, Tatsuya²  Pedersen, Lars C.²  Negishi, Masahiko² 

[1] Department of Chemistry, University of North Carolina, Chapel Hill, NC 27541, USA;Pharmacogenetics Section, Laboratory of Reproductive and Developmental Toxicology, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC 27709, USA

关键词: Lysine-614;    Sulfotransferase;   

DOI  :  10.1016/S0014-5793(98)00913-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

An active sulfotransferase (ST, residues 558–882) domain of the human heparan sulfate N-deacetylase/N-sulfotransferase (hHSNST) has been identified by aligning the amino acid sequence of hHSNST to that of mouse estrogen sulfotransferase (EST). The bacterially expressed ST domain transfers the 5′-sulfuryl group of 3′-phosphoadenosine-5′-phosphosulfate (PAPS) to only deacetylated heparin with an efficiency similar to that previously reported for the purified rat HSNST. Moreover, the K _{m PAPS} (2.1 μM) of the ST domain is also similar to that of the rat enzyme. Lys⁴⁸ is a key residue in mEST catalysis. The residue corresponding to Lys⁴⁸ is conserved in all known heparan sulfate sulfotransferases (Lys⁶¹⁴ in the ST domain of hHSNST). Mutation of Lys⁶¹⁴ to Ala abolishes N-sulfotransferase activity, indicating an important catalytic role of Lys⁶¹⁴ in the ST domain. Crystals of the ST domain have been grown (orthorhombic space group P2₁2₁2) with diffraction to 2.5 A resolution.

【 授权许可】

Unknown