Structure and protein-protein interactions of methanol dehydrogenase from Methylococcus capsulatus (Bath).
Structure and protein-protein interactions of methanol dehydrogenase from Methylococcus capsulatus (Bath).
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DOI：
10.1021/bi500850j
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发表时间：
2014-10-07
期刊：
影响因子：
2.9
通讯作者：
Rosenzweig, Amy C.
中科院分区：
文献类型：
--
作者：
Culpepper, Megen A.;Rosenzweig, Amy C.
在其代谢途径的初始阶段，甲烷营养菌利用甲烷单加氧酶（MMOs）将甲烷氧化为甲醇，利用甲醇脱氢酶（MDHs）将甲醇氧化为甲醛。一些证据表明，膜结合或颗粒MMO （pMMO）和MDH相互作用形成代谢超复合体。为了进一步研究这种超复合物的存在可能性，我们对来自荚膜甲基球菌（Methylococcus capsulatus, Bath）的天然MDH进行了纯化，并用多角度光散射和x射线晶体学对其进行了尺寸排斥层析和表征。M. capsulatus (Bath) MDH在溶液中主要是二聚体，尽管在较高的蛋白质浓度下会形成分子质量约450 - 560kda的低聚物种。分辨率为2.57 Å的晶体结构显示出与其他MDH结构相似的整体褶皱和α2β2二聚体结构。此外，生物层干涉研究表明MDH与M. capsulatus (Bath) pMMO之间以及MDH与M. capsulatus (Bath) pMMO （spmoB）的截断重组质周结构域之间存在特异性蛋白-蛋白相互作用。KD值分别为833±409 nM和9.0±7.7 μM。生化数据结合对MDH结构中观察到的晶格相互作用的分析表明，MDH和pMMO不是作为一个离散的化学计量复合物，而是作为一个由胞质内膜支撑的更大的组装体。
In the initial steps of their metabolic pathway, methanotrophic bacteria oxidize methane to methanol with methane monooxygenases (MMOs) and methanol to formaldehyde with methanol dehydrogenases (MDHs). Several lines of evidence suggest that the membrane-bound or particulate MMO (pMMO) and MDH interact to form a metabolic supercomplex. To further investigate the possible existence of such a supercomplex, native MDH from Methylococcus capsulatus (Bath) has been purified and characterized by size exclusion chromatography with multi-angle light scattering and X-ray crystallography. M. capsulatus (Bath) MDH is primarily a dimer in solution, although an oligomeric species with a molecular mass of ∼450–560 kDa forms at higher protein concentrations. The 2.57 Å resolution crystal structure reveals an overall fold and α2β2 dimeric architecture similar to those of other MDH structures. In addition, biolayer interferometry studies demonstrate specific protein–protein interactions between MDH and M. capsulatus (Bath) pMMO as well as between MDH and the truncated recombinant periplasmic domains of M. capsulatus (Bath) pMMO (spmoB). These interactions exhibit KD values of 833 ± 409 nM and 9.0 ± 7.7 μM, respectively. The biochemical data combined with analysis of the crystal lattice interactions observed in the MDH structure suggest a model in which MDH and pMMO associate not as a discrete, stoichiometric complex but as a larger assembly scaffolded by the intracytoplasmic membranes.
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DOI：
10.1107/s090744491003982x
发表时间：
2011-04
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Evans PR
通讯作者：
Evans PR
影响因子：
64.8
作者：
通讯作者：
--
影响因子：
4.4
作者：
FASSEL, TA;BUCHHOLZ, LA;REMSEN, CC
通讯作者：
REMSEN, CC
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
DOI：
10.1099/00221287-143-5-1729
发表时间：
1997-05-01
期刊：
MICROBIOLOGY-UK
影响因子：
--
作者：
Chistoserdova, L;Lidstrom, ME
通讯作者：
Lidstrom, ME