Recognition of sulfation pattern of chondroitin sulfate by uronosyl 2-O-sulfotransferase
Recognition of sulfation pattern of chondroitin sulfate by uronosyl 2-O-sulfotransferase
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DOI：
10.1074/jbc.m508816200
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发表时间：
2005-11-25
影响因子：
4.8
通讯作者：
Habuchi, O
中科院分区：
文献类型：
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作者：
Ohtake, S;Kimata, K;Habuchi, O
我们先前已经证明，在硫酸软骨素（CS）的非还原末端存在高度硫酸化的序列GalNAc（4，6-SO 4）-GlcA（2SO（4））-GalNAc（6SO（4）），并且该结构是由独特的序列GalNAc（4SO（4））-GlcA（2SO（4））GalNAc（6SO（4））通过N-乙酰半乳糖胺4-硫酸6-O-磺基转移酶硫酸化合成的。尿糖基2-O-磺基转移酶（2 OST）将硫酸根从3 '-磷酸腺苷5'-磷酸硫酸盐（PAPS）转移到CS的GlcA残基的2位，预期参与这些结构的合成;然而，2 OST关于受体硫酸化模式识别的特异性在很大程度上仍不清楚。在本研究中，我们研究了2 OST在识别靶向GlcA残基周围硫酸化模式方面的特异性。重组2 OST可硫酸化CS-A、CS-C和硫酸化皮肤素。当与重组2 OST和[S-35] PAPS反应后由CS-A形成的[S-35]糖胺聚糖用软骨素酶ACII进行有限消化时，得到放射性四糖（Tetra A）作为唯一的中间产物。通过酶促反应和化学反应，确定Tetra A的序列为Delta HexA-GalNAc（4SO（4））GlcA（2SO（4））-GalNAc（6SO（4））。这些观察结果表明，2 OST优先将硫酸盐转移到位于独特序列-GalNAc（4SO（4））-GlcA-GalNAc（6SO（4））-中的GlcA残基。以不同硫酸化模式的寡糖为受体时，三糖和四糖中，GalNAc（4SO（4））-GlcA-GalNAc（6SO（4））和GlcA-GalNAc（4SO（4））GlcA-GalNAc（6SO（4））分别是2 OST的最佳受体。这些结果表明，2 OST可能参与CS-A中发现的高度硫酸化结构的合成。
We have shown previously that a highly sulfated sequence, GalNAc( 4,6-SO4)-GlcA(2SO(4))-GalNAc(6SO(4)), is present at the nonreducing terminal of chondroitin sulfate (CS), and this structure was synthesized from a unique sequence, GalNAc(4SO(4))-GlcA(2SO(4))GalNAc( 6SO(4)), by sulfation with N-acetylgalactosamine 4-sulfate 6-O-sulfotransferase. Uronosyl 2-O-sulfotrasferase (2OST), which transfers sulfate from 3'-phosphoadenosine 5'-phosphosulfate ( PAPS) to position 2 of the GlcA residue of CS, is expected to be involved in synthesis of these structures; however, the specificity of 2OST concerning recognition of the sulfation pattern of the acceptor has largely remained unclear. In the present study, we examined the specificity of 2OST in terms of recognition of the sulfation pattern around the targeting GlcA residue. The recombinant 2OST could sulfate CS-A, CS-C, and desulfated dermatan sulfate. When [S-35] glycosaminoglycans formed from CS-A after the reaction with the recombinant 2OST and [S-35] PAPS were subjected to limited digestion with chondroitinase ACII, a radioactive tetrasaccharide (Tetra A) was obtained as a sole intermediate product. The sequence of Tetra A was found to be Delta HexA-GalNAc(4SO(4))GlcA(2SO(4))-GalNAc(6SO(4)) by enzymatic and chemical reactions. These observations indicate that 2OST transfers sulfate preferentially to the GlcA residue located in a unique sequence, -GalNAc(4SO(4))-GlcA-GalNAc(6SO(4))-. When oligosaccharides with different sulfation patterns were used as the acceptor, GalNAc(4SO(4))-GlcA-GalNAc(6SO(4)) and GlcA-GalNAc(4SO(4))GlcA-GalNAc( 6SO(4)) were the best acceptors for 2OST among trisaccharides and tetrasaccharides, respectively. These results suggest that 2OST may be involved in the synthesis of the highly sulfated structure found in CS-A.