• 文献检索
  • 文档翻译
  • 深度研究
  • 学术资讯
  • Suppr Zotero 插件Zotero 插件
  • 邀请有礼
  • 套餐&价格
  • 历史记录
应用&插件
Suppr Zotero 插件Zotero 插件浏览器插件Mac 客户端Windows 客户端微信小程序
定价
高级版会员购买积分包购买API积分包
服务
文献检索文档翻译深度研究API 文档MCP 服务
关于我们
关于 Suppr公司介绍联系我们用户协议隐私条款
关注我们

Suppr 超能文献

核心技术专利：CN118964589B侵权必究
粤ICP备2023148730 号-1Suppr @ 2026

文献检索

告别复杂PubMed语法，用中文像聊天一样搜索，搜遍4000万医学文献。AI智能推荐，让科研检索更轻松。

立即免费搜索

文件翻译

保留排版，准确专业，支持PDF/Word/PPT等文件格式，支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述，25分钟生成高质量综述，智能提取关键信息，辅助科研写作。

立即免费体验

天然固氮控制蛋白NifA和NifL的纯化及体外活性

Purification and in vitro activities of the native nitrogen fixation control proteins NifA and NifL.

作者信息

Austin S, Buck M, Cannon W, Eydmann T, Dixon R

机构信息

Nitrogen Fixation Laboratory, University of Sussex, Brighton, England.

出版信息

J Bacteriol. 1994 Jun;176(12):3460-5. doi: 10.1128/jb.176.12.3460-3465.1994.

DOI:10.1128/jb.176.12.3460-3465.1994
PMID:8206822
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC205532/
Abstract

The prokaryotic enhancer-binding protein NifA stimulates transcription at a distance by binding to sequences upstream of nitrogen fixation (nif) promoters and catalyzing the formation of open promoter complexes by RNA polymerase containing the alternative sigma factor, sigma 54. The activity of NifA in vivo is modulated by the negative regulatory protein NifL in response to environmental oxygen and fixed nitrogen. To date, a detailed biochemical analysis of these proteins from the model diazotroph Klebsiella pneumoniae has been hindered by their insolubility. We have now purified NifA and NifL from Azotobacter vinelandii in their native form. NifA is competent in specific DNA binding, transcriptional activation, and response to negative regulation by NifL in vitro. In contrast to the conserved mechanism of phosphotransfer demonstrated by other two-component regulatory systems, our results support a model in which NifL regulates the activity of NifA via a protein-protein steric block interaction rather than a catalytic modification of NifA.

摘要

原核生物增强子结合蛋白NifA通过与固氮（nif）启动子上游的序列结合，并催化含有替代sigma因子sigma 54的RNA聚合酶形成开放启动子复合物，从而在远距离刺激转录。NifA在体内的活性受负调控蛋白NifL的调节，以响应环境中的氧气和固定氮。迄今为止，来自模式固氮菌肺炎克雷伯菌的这些蛋白质的详细生化分析因它们的不溶性而受阻。我们现在已经从维涅兰德固氮菌中以天然形式纯化了NifA和NifL。NifA在体外具有特异性DNA结合、转录激活以及对NifL负调控的响应能力。与其他双组分调节系统所展示的保守磷酸转移机制不同，我们的结果支持一种模型，即NifL通过蛋白质-蛋白质空间位阻相互作用而非对NifA的催化修饰来调节NifA的活性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c28f/205532/6e58cb327cee/jbacter00030-0032-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c28f/205532/3fa48b9c6624/jbacter00030-0029-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c28f/205532/fb79f23b3b72/jbacter00030-0029-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c28f/205532/d9c9c5c26248/jbacter00030-0030-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c28f/205532/64dddaa09d4f/jbacter00030-0031-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c28f/205532/6e58cb327cee/jbacter00030-0032-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c28f/205532/3fa48b9c6624/jbacter00030-0029-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c28f/205532/fb79f23b3b72/jbacter00030-0029-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c28f/205532/d9c9c5c26248/jbacter00030-0030-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c28f/205532/64dddaa09d4f/jbacter00030-0031-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c28f/205532/6e58cb327cee/jbacter00030-0032-a.jpg

相似文献

1
Purification and in vitro activities of the native nitrogen fixation control proteins NifA and NifL.天然固氮控制蛋白NifA和NifL的纯化及体外活性
J Bacteriol. 1994 Jun;176(12):3460-5. doi: 10.1128/jb.176.12.3460-3465.1994.
2
Regulation of nitrogen fixation in Klebsiella pneumoniae and Azotobacter vinelandii: NifL, transducing two environmental signals to the nif transcriptional activator NifA.肺炎克雷伯菌和棕色固氮菌中固氮作用的调控：NifL，将两种环境信号传递给固氮转录激活因子NifA。
J Mol Microbiol Biotechnol. 2002 May;4(3):235-42.
3
Transcriptional activation of the nitrogenase promoter in vitro: adenosine nucleotides are required for inhibition of NIFA activity by NIFL.体外固氮酶启动子的转录激活：腺苷核苷酸是NIFL抑制NIFA活性所必需的。
J Bacteriol. 1995 Mar;177(5):1186-95. doi: 10.1128/jb.177.5.1186-1195.1995.
4
Isolation and properties of the complex between the enhancer binding protein NIFA and the sensor NIFL.增强子结合蛋白NIFA与传感蛋白NIFL之间复合物的分离及特性
J Bacteriol. 1999 Aug;181(15):4461-8. doi: 10.1128/JB.181.15.4461-4468.1999.
5
The C-terminal domain of NifL is sufficient to inhibit NifA activity.固氮调节蛋白L（NifL）的C末端结构域足以抑制固氮激活蛋白（NifA）的活性。
J Bacteriol. 1995 Sep;177(17):5078-87. doi: 10.1128/jb.177.17.5078-5087.1995.
6
Role of Escherichia coli nitrogen regulatory genes in the nitrogen response of the Azotobacter vinelandii NifL-NifA complex.大肠杆菌氮调节基因在棕色固氮菌NifL-NifA复合物氮响应中的作用。
J Bacteriol. 2001 May;183(10):3076-82. doi: 10.1128/JB.183.10.3076-3082.2001.
7
Role of the amino-terminal GAF domain of the NifA activator in controlling the response to the antiactivator protein NifL.固氮激活蛋白NifA的氨基端GAF结构域在调控对抗激活蛋白NifL反应中的作用
Mol Microbiol. 2004 Jun;52(6):1731-44. doi: 10.1111/j.1365-2958.2004.04089.x.
8
In vitro activity of NifL, a signal transduction protein for biological nitrogen fixation.NifL的体外活性，一种生物固氮的信号转导蛋白。
J Bacteriol. 1993 Dec;175(23):7683-8. doi: 10.1128/jb.175.23.7683-7688.1993.
9
Iron is required to relieve inhibitory effects on NifL on transcriptional activation by NifA in Klebsiella pneumoniae.肺炎克雷伯菌中，需要铁来解除NifL对NifA转录激活的抑制作用。
J Bacteriol. 1996 Aug;178(15):4679-87. doi: 10.1128/jb.178.15.4679-4687.1996.
10
Concerted inhibition of the transcriptional activation functions of the enhancer-binding protein NIFA by the anti-activator NIFL.抗激活因子NIFL对增强子结合蛋白NIFA转录激活功能的协同抑制作用。
Mol Microbiol. 2001 Jan;39(2):480-93. doi: 10.1046/j.1365-2958.2001.02243.x.

引用本文的文献

1
Nitrogen-Fixing Gamma Proteobacteria -A Blueprint for Nitrogen-Fixing Plants?固氮γ-变形菌——固氮植物的蓝本？
Microorganisms. 2024 Oct 18;12(10):2087. doi: 10.3390/microorganisms12102087.
2
Structural insights into redox signal transduction mechanisms in the control of nitrogen fixation by the NifLA system.结构洞察氮固定的 NifLA 系统控制中的氧化还原信号转导机制。
Proc Natl Acad Sci U S A. 2023 Jul 25;120(30):e2302732120. doi: 10.1073/pnas.2302732120. Epub 2023 Jul 17.
3
Molecular Mechanism and Agricultural Application of the NifA-NifL System for Nitrogen Fixation.

本文引用的文献

1
The Azotobacter vinelandii nifL-like gene: nucleotide sequence analysis and regulation of expression.棕色固氮菌的类nifL基因：核苷酸序列分析及表达调控
Mol Gen Genet. 1993 Mar;237(3):400-6. doi: 10.1007/BF00279444.
2
The Klebsiella pneumoniae nifJ promoter: analysis of promoter elements regulating activation by the NifA promoter.肺炎克雷伯菌nifJ启动子：调控NifA启动子激活的启动子元件分析
Mol Microbiol. 1993 Mar;7(6):1007-21. doi: 10.1111/j.1365-2958.1993.tb01192.x.
3
Activity of purified NIFA, a transcriptional activator of nitrogen fixation genes.
NifA-NifL 系统固氮的分子机制及其在农业中的应用。
Int J Mol Sci. 2023 Jan 4;24(2):907. doi: 10.3390/ijms24020907.
4
Comparative genomic analyses of the bacterial phosphotransferase system.细菌磷酸转移酶系统的比较基因组分析
Microbiol Mol Biol Rev. 2005 Dec;69(4):608-34. doi: 10.1128/MMBR.69.4.608-634.2005.
5
A crucial arginine residue is required for a conformational switch in NifL to regulate nitrogen fixation in Azotobacter vinelandii.一个关键的精氨酸残基是棕色固氮菌中NifL构象转换以调节固氮作用所必需的。
Proc Natl Acad Sci U S A. 2004 Nov 16;101(46):16316-21. doi: 10.1073/pnas.0405312101. Epub 2004 Nov 8.
6
DNA binding activity of the Escherichia coli nitric oxide sensor NorR suggests a conserved target sequence in diverse proteobacteria.大肠杆菌一氧化氮传感器NorR的DNA结合活性表明，不同变形菌中存在保守的靶序列。
J Bacteriol. 2004 Oct;186(19):6656-60. doi: 10.1128/JB.186.19.6656-6660.2004.
7
The NifL-NifA System: a multidomain transcriptional regulatory complex that integrates environmental signals.NifL-NifA系统：一种整合环境信号的多结构域转录调控复合体。
J Bacteriol. 2004 Feb;186(3):601-10. doi: 10.1128/JB.186.3.601-610.2004.
8
Mutant forms of the Azotobacter vinelandii transcriptional activator NifA resistant to inhibition by the NifL regulatory protein.对固氮螺菌属维涅兰德氏菌转录激活因子NifA的突变形式具有抗性，该突变形式不受NifL调节蛋白抑制。
J Bacteriol. 2002 Dec;184(24):6777-85. doi: 10.1128/JB.184.24.6777-6785.2002.
9
An active role for a structured B-linker in effector control of the sigma54-dependent regulator DmpR.结构化B连接体在σ54依赖性调节因子DmpR的效应控制中发挥积极作用。
EMBO J. 2001 Feb 15;20(4):819-27. doi: 10.1093/emboj/20.4.819.
10
Protein-protein interactions in the complex between the enhancer binding protein NIFA and the sensor NIFL from Azotobacter vinelandii.来自棕色固氮菌的增强子结合蛋白NIFA与传感器NIFL复合物中的蛋白质-蛋白质相互作用。
J Bacteriol. 2001 Feb;183(4):1359-68. doi: 10.1128/JB.183.4.1359-1368.2001.
纯化的NIFA（一种固氮基因的转录激活因子）的活性。
Proc Natl Acad Sci U S A. 1993 Mar 15;90(6):2266-70. doi: 10.1073/pnas.90.6.2266.
4
The sigma 54 bacterial enhancer-binding protein family: mechanism of action and phylogenetic relationship of their functional domains.σ54细菌增强子结合蛋白家族：其功能域的作用机制及系统发育关系
J Bacteriol. 1993 Oct;175(19):6067-74. doi: 10.1128/jb.175.19.6067-6074.1993.
5
Sequence and molecular analysis of the nifL gene of Azotobacter vinelandii.棕色固氮菌nifL基因的序列与分子分析
Mol Microbiol. 1993 Aug;9(4):869-79. doi: 10.1111/j.1365-2958.1993.tb01745.x.
6
DNA distortion and nucleation of local DNA unwinding within sigma-54 (sigma N) holoenzyme closed promoter complexes.σ⁵⁴（σᴺ）全酶封闭启动子复合物内的DNA扭曲及局部DNA解旋的成核作用。
J Biol Chem. 1994 Apr 15;269(15):11563-71.
7
Defining a bacteriophage T4 late promoter: absence of a "-35" region.界定噬菌体T4晚期启动子：不存在“-35”区域。
Cell. 1984 Jan;36(1):211-9. doi: 10.1016/0092-8674(84)90091-6.
8
Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes.利用噬菌体T7 RNA聚合酶指导克隆基因的选择性高水平表达。
J Mol Biol. 1986 May 5;189(1):113-30. doi: 10.1016/0022-2836(86)90385-2.
9
Sequence and domain relationships of ntrC and nifA from Klebsiella pneumoniae: homologies to other regulatory proteins.
EMBO J. 1986 Feb;5(2):441-7. doi: 10.1002/j.1460-2075.1986.tb04230.x.
10
NifA-dependent in vivo protection demonstrates that the upstream activator sequence of nif promoters is a protein binding site.依赖于NifA的体内保护作用表明，固氮酶基因启动子的上游激活序列是一个蛋白质结合位点。
Proc Natl Acad Sci U S A. 1988 Dec;85(24):9401-5. doi: 10.1073/pnas.85.24.9401.