Supramolecular Complex Formation and Crystallization of Isocitrate Dehydrogenase from Thermus thermophilus HB8: Preliminary Studies with X-Ray Crystallography and Atomic Force Microscopy
Supramolecular Complex Formation and Crystallization of Isocitrate Dehydrogenase from Thermus thermophilus HB8: Preliminary Studies with X-Ray Crystallography and Atomic Force Microscopy
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嗜热栖热菌 HB8 异柠檬酸脱氢酶的超分子复合物形成和结晶：X 射线晶体学和原子力显微镜的初步研究
DOI：
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发表时间：
2008
期刊：
影响因子：
--
通讯作者：
K. Miyazaki
中科院分区：
文献类型：
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作者：
N. Ishii;K. Umemura;K. Miyazaki
利用原子力显微镜（AFM）对嗜热真细菌Thermus thermophilus HB 8的热稳定异柠檬酸脱氢酶（ICDH）晶体表面进行了观察，结果表明该晶体由巨大的同质复合椭球体组成，长轴和短轴平均直径分别为18.6 nm和10.9 nm。通过气相扩散法从含有1.4M乙酸钠作为沉淀物的100 mM二甲胂酸钠溶液（pH 6.6-8.4）中获得最长轴上约0.4mm的厚菱形晶体，并以3.7 μ m分辨率衍射X射线。晶体属于单斜晶系，空间群为C2，晶胞尺寸为a=495.5，B=189.2，c=336.2 μ m，β=126.4°，表明一个不对称单元含有超过33个分子，分子量为54.2 kDa。基于X射线方法获得的数据的计算结果与AFM观察结果吻合良好。这些结果表明，T.嗜热菌HB 8 ICDH分子在晶格中一个在另一个之上堆叠作为预先形成的超分子纳米结构。该系统似乎适合于进一步调查使用自下而上的方法与蛋白质的纳米结构的自相关的建设。
Atomic force microscopy (AFM) observation of a crystal surface of the thermostable isocitrate dehydrogenase (ICDH) from a thermophilic eubacterium, Thermus thermophilus HB8, suggested that the crystal consists of huge homo-complexed ellipsoidal bodies of the protein, with averaged long- and short-axis diameters of 18.6 nm and 10.9 nm respectively. Thick diamond-shaped crystals of about 0.4 mm on the longest axis were obtained by the vapor diffusion method from a solution of 100 mM sodium cacodylate, pH 6.6–8.4, containing 1.4 M sodium acetate as the precipitate, and diffracted X-rays at 3.7 Å resolution. The crystals belonged to the monoclinic lattice type with space group C2 and had cell dimensions of a=495.5, b=189.2, c=336.2 Å, and β=126.4°, indicating that an asymmetric unit contained more than 33 molecules with a molecular mass of 54.2 kDa. Calculations based on data obtained by the X-ray method showed good agreement with AFM observation. These results suggest the possibility that the residing T. thermophilus HB8 ICDH molecules are piled one on top another as a preformed supramolecular nano-architecture in the crystal lattice. The system appears suitable for further investigation using a bottom-up approach to the self-associated construction of nano-architectures with proteins.
DOI：
10.1073/pnas.86.22.8635
发表时间：
1989-11-01
影响因子：
11.1
作者：
HURLEY, JH;THORSNESS, PE;STROUD, RM
通讯作者：
STROUD, RM
影响因子：
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作者：
Krieger IV;Freundlich JS;Gawandi VB;Roberts JP;Gawandi VB;Sun Q;Owen JL;Fraile MT;Huss SI;Lavandera JL;Ioerger TR;Sacchettini JC
通讯作者：
Sacchettini JC
影响因子：
56.9
作者：
HURLEY, JH;DEAN, AM;STROUD, RM
通讯作者：
STROUD, RM