赖氨酸614在人硫酸乙酰肝素N-脱乙酰酶/N-磺基转移酶的磺基转移酶活性中的作用。
A role of Lys614 in the sulfotransferase activity of human heparan sulfate N-deacetylase/N-sulfotransferase.
作者信息
Sueyoshi T, Kakuta Y, Pedersen L C, Wall F E, Pedersen L G, Negishi M
机构信息
Pharmacogenetics Section, Laboratory of Reproductive and Developmental Toxicology, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC 27709, USA.
出版信息
FEBS Lett. 1998 Aug 21;433(3):211-4. doi: 10.1016/s0014-5793(98)00913-2.
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 microM) 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 P2(1)2(1)2) with diffraction to 2.5 A resolution.
通过将人硫酸乙酰肝素N - 脱乙酰酶/N - 磺基转移酶（hHSNST）的氨基酸序列与小鼠雌激素磺基转移酶（EST）的序列进行比对，已鉴定出hHSNST的一个活性磺基转移酶（ST，第558 - 882位氨基酸残基）结构域。细菌表达的ST结构域将3'-磷酸腺苷-5'-磷酸硫酸酯（PAPS）的5'-硫酰基仅转移至脱乙酰化的肝素，其效率与先前报道的纯化大鼠HSNST相似。此外，ST结构域的K(m,PAPS)（2.1 microM）也与大鼠酶的相似。Lys48是mEST催化中的关键残基。在所有已知的硫酸乙酰肝素磺基转移酶中，与Lys48对应的残基是保守的（hHSNST的ST结构域中的Lys614）。将Lys614突变为Ala会消除N - 磺基转移酶活性，表明Lys614在ST结构域中具有重要的催化作用。已培养出ST结构域的晶体（正交空间群P2(1)2(1)2），衍射分辨率达到2.5 Å。
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