Structural studies of a surface-entropy reduction mutant of O-GlcNAcase
Structural studies of a surface-entropy reduction mutant of O-GlcNAcase
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O-GlcNAcase 表面熵减少突变体的结构研究
DOI：
10.1107/s2059798318016595
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Males A
中科院分区：
文献类型：
--
作者：
Males A
酶O-GlcNAc酶催化多细胞真核生物中O-GlcNAc共/翻译后修饰的去除。考虑到O-GlcNAc化在tau修饰和稳定性中的密切作用，该酶已引起人们的极大兴趣;人O-GlcNAc酶的小分子抑制剂正在临床评估中用于治疗tau蛋白病。考虑到基于结构和基于机制的O-GlcNAcase抑制剂设计的重要性，试图测试是否可以通过表面诱变获得人酶的不同晶体形式。在表面熵减少的指导下，获得了Glu 602 Ala/Glu 605 Ala变体[在Gly 11-Gln 396/Lys 535-Tyr 715构建体上; Roth et al.（2017），Nature Chem.Biol.13，610-612]，这导致了人酶的新晶体形式。晶体接触的增加稳定了蛋白质的无序区域，使88%的结构能够被建模;野生型构建体只有83%。尽管C-末端的结合与野生型一致，但单体A中的Lys 713结合在与突变相关的单体A的−1亚位点，而B单体的活性位点是空的。这种新的晶体形式为增强浸泡实验提供了机会，这些实验对于理解O-GlcNAcase的结合机制和底物特异性至关重要。
The enzyme O-GlcNAcase catalyses the removal of the O-GlcNAc co/post-translational modification in multicellular eukaryotes. The enzyme has become of acute interest given the intimate role of O-GlcNAcylation in tau modification and stability; small-molecular inhibitors of human O-GlcNAcase are under clinical assessment for the treatment of tauopathies. Given the importance of structure-based and mechanism-based inhibitor design for O-GlcNAcase, it was sought to test whether different crystal forms of the human enzyme could be achieved by surface mutagenesis. Guided by surface-entropy reduction, a Glu602Ala/Glu605Ala variant [on the Gly11–Gln396/Lys535–Tyr715 construct; Roth et al. (2017), Nature Chem. Biol. 13, 610–612] was obtained which led to a new crystal form of the human enzyme. An increase in crystal contacts stabilized disordered regions of the protein, enabling 88% of the structure to be modelled; only 83% was possible for the wild-type construct. Although the binding of the C-terminus was consistent with the wild type, Lys713 in monomer A was bound in the −1 subsite of the symmetry-related monomer A and the active sites of the B monomers were vacant. The new crystal form presents an opportunity for enhanced soaking experiments that are essential to understanding the binding mechanism and substrate specificity of O-GlcNAcase.
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实现中小型学术实验室结晶筛选的合理化：PACT/JCSG 策略。
DOI：
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发表时间：
2005
期刊：
Acta Crystallographica Section D: Biological Crystallography
影响因子：
--
作者：
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通讯作者：
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DOI：
10.1107/s0907444905018822
发表时间：
2005-09-01
影响因子：
2.2
作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
van Aalten DM
DOI：
10.1107/s0907444901003122
发表时间：
2001-05-01
影响因子：
2.2
作者：
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通讯作者：
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DOI：
10.1006/bbrc.1995.2149
发表时间：
1995-08-15
影响因子：
3.1
作者：
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通讯作者：
SCHMITZ, B