How to change the oligomeric state of a circular protein assembly: switch from 11-subunit to 12-subunit TRAP suggests a general mechanism.
How to change the oligomeric state of a circular protein assembly: switch from 11-subunit to 12-subunit TRAP suggests a general mechanism.
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DOI：
10.1371/journal.pone.0025296
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Antson AA
中科院分区：
文献类型：
--
作者：
Chen CS;Smits C;Dodson GG;Shevtsov MB;Merlino N;Gollnick P;Antson AA
许多重要的细胞功能是由多亚基环状蛋白寡聚体完成的，其内部几何结构已经进化以满足功能要求。亚基数量可以说是环状蛋白质组装体的最关键参数，影响组装体的内径和外径，并且经常影响蛋白质的功能。虽然已经获得了几个环状蛋白质的准确结构信息，但缺乏关于替代寡聚状态的准确信息阻止了工程化这种转变。在这项研究中，我们使用的细菌转录调节TRAP作为模型系统，以调查的功能，定义的寡聚状态的环状蛋白质，并质疑如何可以操纵的亚基数量。我们发现，虽然枯草芽孢杆菌和嗜热脂肪芽孢杆菌TRAP形成11-亚基寡聚体，耐盐芽孢杆菌TRAP专门形成12-亚基组件。值得注意的是，这两种状态的陷阱是由一个简单的刚体旋转的各个亚基之间的亚基轴周围。我们测试了这样的旋转是否可以诱导插入或缺失突变的亚基接口。使用野生型11-亚基TRAP，我们表明，5个C-末端残基的去除在外侧的亚基间轴或延伸的氨基酸侧链在相反的，内侧，增加了亚基的数量从11到12。我们的研究结果支持TRAP低聚物的晶体结构和本地质谱数据。TRAP寡聚体的亚基数目可以通过在亚基界面处引入缺失或添加突变来操纵。对替代低聚状态的可用和新出现的结构数据的分析表明，相同的原理也可适用于其他环状组装体的亚基数目，这表明缺失/添加方法通常可用于工程化不同低聚状态之间的转换。
Many critical cellular functions are performed by multisubunit circular protein oligomers whose internal geometry has evolved to meet functional requirements. The subunit number is arguably the most critical parameter of a circular protein assembly, affecting the internal and external diameters of the assembly and often impacting on the protein's function. Although accurate structural information has been obtained for several circular proteins, a lack of accurate information on alternative oligomeric states has prevented engineering such transitions. In this study we used the bacterial transcription regulator TRAP as a model system to investigate the features that define the oligomeric state of a circular protein and to question how the subunit number could be manipulated. We find that while Bacillus subtilis and Bacillus stearothermophilus TRAP form 11-subunit oligomers, the Bacillus halodurans TRAP exclusively forms 12-subunit assemblies. Significantly, the two states of TRAP are related by a simple rigid body rotation of individual subunits around inter-subunit axes. We tested if such a rotation could be induced by insertion or deletion mutations at the subunit interface. Using wild type 11-subunit TRAP, we demonstrate that removal of five C-terminal residues at the outer side of the inter-subunit axis or extension of an amino acid side chain at the opposite, inner side, increased the subunit number from 11 to 12. Our findings are supported by crystal structures of TRAP oligomers and by native mass spectrometry data. The subunit number of the TRAP oligomer can be manipulated by introducing deletion or addition mutations at the subunit interface. An analysis of available and emerging structural data on alternative oligomeric states indicates that the same principles may also apply to the subunit number of other circular assemblies suggesting that the deletion/addition approach could be used generally to engineer transitions between different oligomeric states.
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DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
5.6
作者：
Chen, XP;Antson, AA;Gollnick, P
通讯作者：
Gollnick, P
影响因子：
64.8
作者：
Antson, AA;Dodson, EJ;Gollnick, P
通讯作者：
Gollnick, P
影响因子：
15
作者：
McCammon, MG;Hernández, H;Robinson, CV
通讯作者：
Robinson, CV
影响因子：
5.7
作者：
Heddle, Jonathan G.;Yokoyama, Takeshi;Tame, Jeremy R. H.
通讯作者：
Tame, Jeremy R. H.