Topological Analysis of the Membrane-localized Redox-responsive Sensor Kinase PrrB from Rhodobacter sphaeroides 2.4.1*

Topological Analysis of the Membrane-localized Redox-responsive Sensor Kinase PrrB from Rhodobacter sphaeroides 2.4.1*
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来自球形红杆菌 2.4.1* 的膜定位氧化还原响应传感器激酶 PrrB 的拓扑分析

DOI:
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发表时间:
1999
影响因子:
4.8
通讯作者:
S. Kaplan
S. Kaplan
中科院分区:
生物学2区
文献类型:
--
作者:
S. Ouchane;S. Kaplan

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球形红杆菌光合作用基因的表达部分受由膜结合的传感蛋白(PrrB)和反应调节蛋白(PrrA)组成的双组分(PRR)调控系统的调控。基于亲水性图谱的计算机分析预测,PrrB多肽在其氨基末端可能含有6个跨膜结构域,在细胞质中的羧基末端可能含有一个亲水的羧基末端。通过与大肠杆菌胞质定位的碱性磷酸酶和胞质β-半乳糖苷酶的一系列基因融合,研究了PrrB传感器激酶的定位和拓扑结构。构建了18个prrB-PhoA和5个prrB-lacZ融合基因,并在两者中进行了表达。Coli和球形葡萄球菌(R.sphaeroides.)通过酶活性分析和免疫印迹分析来鉴定和定位PrrB在膜上的不同片段。在大肠杆菌中获得的数据通常与在球形葡萄球菌中获得的数据相关联,并支持计算机预测。在理论模型和这些研究结果的基础上,提出了PrrB多肽膜定位的拓扑模型。
Photosynthesis gene expression inRhodobacter sphaeroides is controlled in part by the two-component (Prr) regulatory system composed of a membrane-bound sensor kinase (PrrB) and a response regulator (PrrA). Hydropathy profile-based computer analysis predicted that the PrrB polypeptide could contain six membrane-spanning domains at its amino terminus and a hydrophilic, cytoplasmic carboxyl terminus. Both the localization and the topology of the PrrB sensor kinase have been studied by generating a series of gene fusions with the Escherichia coliperiplasmically localized alkaline phosphatase and the cytoplasmic β-galactosidase. Eighteen prrB-phoA and fiveprrB-lacZ fusions were constructed and expressed in bothE. coli and R. sphaeroides. Enzymatic activity assays and immunoblot analyses were performed to identify and to localize the different segments of PrrB in the membrane. The data obtained in E. coli generally correlated with the data obtained in R. sphaeroides and support the computer predictions. On the basis of the theoretical model and the results provided by these studies, a topological model for the membrane localization of the PrrB polypeptide is proposed.
DOI: --
发表时间: 1991
期刊: The Journal of biological chemistry
影响因子: --
作者:
Yun,CH;VanDoren,SR;Crofts,AR;Gennis,RB
通讯作者: Gennis,RB
DOI: --
发表时间: 1994-04
期刊: BioTechniques
影响因子: 2.7
作者:
Michael E. Kovach;R. Phillips;P. Elzer;R. Roop;K. Peterson
通讯作者: Michael E. Kovach;R. Phillips;P. Elzer;R. Roop;K. Peterson
DOI: --
发表时间: 1992
期刊: The Journal of biological chemistry
影响因子: --
作者:
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通讯作者: Ames,GF
大肠杆菌前脂蛋白信号肽酶(信号肽酶 II)的膜拓扑。
DOI: --
发表时间: 1991
期刊: The Journal of biological chemistry
影响因子: --
作者:
Muñoa,FJ;Miller,KW;Beers,R;Graham,M;Wu,HC
通讯作者: Wu,HC