Subunit Interactions within the Carbon-Phosphorus Lyase Complex from Escherichia coli.
Subunit Interactions within the Carbon-Phosphorus Lyase Complex from Escherichia coli.
复制标题
大肠杆菌碳磷裂解酶复合物内的亚基相互作用。
DOI：
10.1021/acs.biochem.5b00194
复制
发表时间：
2015
期刊：
影响因子：
2.9
通讯作者：
Raushel,FrankM
中科院分区：
文献类型：
--
作者：
Ren,Zhongjie;Ranganathan,Soumya;Zinnel,NathanaelF;Russell,WilliamK;Russell,DavidH;Raushel,FrankM
膦酸酯是一大类具有特征性碳-磷键的有机磷化合物。革兰氏阴性菌中负责膦酸盐利用的基因排列在14个基因的操纵子中。由基因phnGHIJKLM编码的碳-磷裂解酶复合物催化有机膦酸酯的稳定碳-磷键裂解为相应的烃和无机磷酸盐。最近，在大肠杆菌中表达后，纯化了含有五个亚基（PhnG-H-I-J-K）、四个亚基（PhnG-H-I-J）和两个亚基（PhnG-I）的该酶的复合物（Proc.Natl. Acad.科学，联合S. A. 2011，108，11393）。在这里，我们证明了使用质谱，超离心，和化学交联实验，这些复合物是从PhnG 2 I2核心，进一步阐述了添加两个拷贝的PhnH和PhnJ生成PhnG 2 H2 I2 J2。该复合物添加了PhnK的额外亚基以形成PhnG 2 H2 I2 J2 K。五组分复合物的化学交联表明PhnJ与PhnG和PhnI物理相互作用。我们无法通过化学交联证明PhnH或PhnK与任何其他亚基的相互作用。氢-氘交换被用来探测各种复合物内各个亚基的动态特性的改变。与单独的PhnG相比，PhnG的重要区域变得不太容易从PhnG 2 I2复合物内的溶剂中进行氢/氘交换。在PhnG 2 I2和PhnG 2 H2 I2 J2 K中，PhnI的特定区域在H/D交换率方面表现出显著差异。
Phosphonates are a large class of organophosphorus compounds with a characteristic carbon–phosphorus bond. The genes responsible for phosphonate utilization in Gram-negative bacteria are arranged in an operon of 14 genes. The carbon–phosphorus lyase complex, encoded by the genesphnGHIJKLM, catalyzes the cleavage of the stable carbon–phosphorus bond of organophosphonates to the corresponding hydrocarbon and inorganic phosphate. Recently, complexes of this enzyme containing five subunits (PhnG-H-I-J-K), four subunits (PhnG-H-I-J), and two subunits (PhnG-I) were purified after expression inEscherichia coli(Proc. Natl. Acad. Sci., U. S. A. 2011, 108, 11393). Here we demonstrated using mass spectrometry, ultracentrifugation, and chemical cross-linking experiments that these complexes are formed from a PhnG2I2core that is further elaborated by the addition of two copies each of PhnH and PhnJ to generate PhnG2H2I2J2. This complex adds an additional subunit of PhnK to form PhnG2H2I2J2K. Chemical cross-linking of the five-component complex demonstrated that PhnJ physically interacts with both PhnG and PhnI. We were unable to demonstrate the interaction of PhnH or PhnK with any other subunits by chemical cross-linking. Hydrogen–deuterium exchange was utilized to probe for alterations in the dynamic properties of individual subunits within the various complexes. Significant regions of PhnG become less accessible to hydrogen/deuterium exchange from solvent within the PhnG2I2complex compared with PhnG alone. Specific regions of PhnI exhibited significant differences in the H/D exchange rates in PhnG2I2and PhnG2H2I2J2K.
登录
查看更多内容
DOI：
--
发表时间：
1985-03
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
G. Grynkiewicz;M. Poenie;Roger Y. TsienB
通讯作者：
G. Grynkiewicz;M. Poenie;Roger Y. TsienB
DOI：
10.1083/jcb.98.2.717
发表时间：
1984-02
期刊：
The Journal of cell biology
影响因子：
--
作者：
Tanasugarn L;McNeil P;Reynolds GT;Taylor DL
通讯作者：
Taylor DL
影响因子：
4.1
作者：
T. Hallam;A. Sanchez;T. Rink
通讯作者：
T. Rink
使用捕获指示剂测量和操作胞质 Ca2。
DOI：
--
发表时间：
1984
期刊：
Kroc Foundation series
影响因子：
--
作者：
Tsien,RY
通讯作者：
Tsien,RY