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A role of Lys 614 in the sulfotransferase activity of human heparan sulfate N ‐deacetylase/ N ‐sulfotransferase
Author(s) -
Sueyoshi Tatsuya,
Kakuta Yoshimitsu,
Pedersen Lars C.,
Wall Frances E.,
Pedersen Lee G.,
Negishi Masahiko
Publication year - 1998
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(98)00913-2
Subject(s) - sulfotransferase , chemistry , biochemistry , heparan sulfate , sulfation , stereochemistry , residue (chemistry) , enzyme , heparin
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. Lys 48 is a key residue in mEST catalysis. The residue corresponding to Lys 48 is conserved in all known heparan sulfate sulfotransferases (Lys 614 in the ST domain of hHSNST). Mutation of Lys 614 to Ala abolishes N ‐sulfotransferase activity, indicating an important catalytic role of Lys 614 in the ST domain. Crystals of the ST domain have been grown (orthorhombic space group P2 1 2 1 2) with diffraction to 2.5 A resolution.