Use of cyclic peptides to induce crystallization: case study with prolyl hydroxylase domain 2.
Use of cyclic peptides to induce crystallization: case study with prolyl hydroxylase domain 2.
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使用环肽诱导结晶：脯氨酰羟化酶结构域 2 的案例研究。
DOI：
10.1038/s41598-020-76307-8
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发表时间：
2020-12-15
影响因子：
4.6
通讯作者：
Schofield CJ
中科院分区：
文献类型：
--
作者：
Chowdhury R;Abboud MI;McAllister TE;Banerji B;Bhushan B;Sorensen JL;Kawamura A;Schofield CJ
结晶是大分子晶体学中的瓶颈;即使当蛋白质结晶时，晶体堆积也经常影响配体结合和蛋白质-蛋白质相互作用界面，这是功能和药物发现研究的关键点。人缺氧诱导因子脯氨酰羟化酶2（PHD 2）容易结晶为同源三聚体，但具有空间阻断的活性位点。我们探索了旨在通过蛋白质修饰和在其活性位点和其他地方结合的分子来改变PHD 2晶体包装的策略。在观察到尽管在溶液中弱抑制/结合，但琥珀酰胺酸衍生物容易使PHD 2结晶之后，我们探索了在没有活性位点结合的情况下诱导结晶的方法。通过mRNA展示获得的环肽与PHD 2紧密结合，远离活性位点。它们有效地使PHD 2以不同形式结晶，无论有/没有底物，显然是通过促进涉及结合到C-末端区域的寡聚化。虽然我们的工作涉及一个特定的案例研究，与其他人一起，结果表明，mRNA展示衍生的环肽可能是有用的，在具有挑战性的蛋白质结晶的情况下。
Crystallization is the bottleneck in macromolecular crystallography; even when a protein crystallises, crystal packing often influences ligand-binding and protein–protein interaction interfaces, which are the key points of interest for functional and drug discovery studies. The human hypoxia-inducible factor prolyl hydroxylase 2 (PHD2) readily crystallises as a homotrimer, but with a sterically blocked active site. We explored strategies aimed at altering PHD2 crystal packing by protein modification and molecules that bind at its active site and elsewhere. Following the observation that, despite weak inhibition/binding in solution, succinamic acid derivatives readily enable PHD2 crystallization, we explored methods to induce crystallization without active site binding. Cyclic peptides obtained via mRNA display bind PHD2 tightly away from the active site. They efficiently enable PHD2 crystallization in different forms, both with/without substrates, apparently by promoting oligomerization involving binding to the C-terminal region. Although our work involves a specific case study, together with those of others, the results suggest that mRNA display-derived cyclic peptides may be useful in challenging protein crystallization cases.
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影响因子：
4.8
作者：
Elkins, JM;Hewitson, KS;Schofield, CJ
通讯作者：
Schofield, CJ
影响因子：
64.8
作者：
Han, Zhifu;Niu, Tianhui;Chai, Jijie
通讯作者：
Chai, Jijie
影响因子：
56.9
作者：
Jaakkola, P;Mole, DR;Ratcliffe, PJ
通讯作者：
Ratcliffe, PJ
影响因子：
16.6
作者：
Kawamura A;Münzel M;Kojima T;Yapp C;Bhushan B;Goto Y;Tumber A;Katoh T;King ON;Passioura T;Walport LJ;Hatch SB;Madden S;Müller S;Brennan PE;Chowdhury R;Hopkinson RJ;Suga H;Schofield CJ
通讯作者：
Schofield CJ
影响因子：
5.7
作者：
Horita, Shoichiro;Scotti, John S.;Thinnes, Cyrille;Mottaghi-Taromsari, Yousef S.;Thalhammer, Armin;Ge, Wei;Aik, WeiShen;Loenarz, Christoph;Schofield, Christopher J.;McDonough, Michael A.
通讯作者：
McDonough, Michael A.