Structural insights into HDAC6 tubulin deacetylation and its selective inhibition
Structural insights into HDAC6 tubulin deacetylation and its selective inhibition
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DOI：
10.1038/nchembio.2140
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发表时间：
2016-09-01
影响因子：
14.8
通讯作者：
Matthias, Patrick
中科院分区：
文献类型：
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作者：
Miyake, Yasuyuki;Keusch, Jeremy J.;Matthias, Patrick
我们报告了斑马鱼组蛋白脱乙酰基酶6（HDAC 6）催化结构域的晶体结构，这些催化结构域串联或作为单一结构域与阿司他丁A（TSA）的（R）和（S）对映体或HDAC 6特异性抑制剂nexturastat A复合。串联结构域与结构域间连接体一起形成具有假双重对称性的椭圆形复合物。我们确定了两个催化结构域之间的重要活性位点差异，并揭示了HDAC6选择性抑制剂的结合模式。用（R）-和（S）-TSA进行的HDAC抑制测定显示，（R）-TSA是宽范围抑制剂，而（S）-TSA对HDAC 6具有中等选择性。我们确定了一个独特的定位α-螺旋和一个灵活的色氨酸残基的环连接α-螺旋H20到H21的生理底物微管蛋白的脱乙酰化的关键。使用单分子测量和生物化学测定，我们证明HDAC 6催化结构域2在微管的管腔中脱乙酰化α-微管蛋白赖氨酸40，但其优选底物是未聚合的微管蛋白。
We report crystal structures of zebrafish histone deacetylase 6 (HDAC6) catalytic domains in tandem or as single domains in complex with the (R) and (S) enantiomers of trichostatin A (TSA) or with the HDAC6-specific inhibitor nexturastat A. The tandem domains formed, together with the inter-domain linker, an ellipsoid-shaped complex with pseudo-twofold symmetry. We identified important active site differences between both catalytic domains and revealed the binding mode of HDAC6 selective inhibitors. HDAC inhibition assays with (R)-and (S)-TSA showed that (R)-TSA was a broad-range inhibitor, whereas (S)-TSA had moderate selectivity for HDAC6. We identified a uniquely positioned a-helix and a flexible tryptophan residue in the loop joining a-helices H20 to H21 as critical for deacetylation of the physiologic substrate tubulin. Using single-molecule measurements and biochemical assays we demonstrated that HDAC6 catalytic domain 2 deacetylated a-tubulin lysine 40 in the lumen of microtubules, but that its preferred substrate was unpolymerized tubulin.