Biochemistry of Fatty Acid Uptake in Escherichia Coli

大肠杆菌摄取脂肪酸的生物化学

基本信息

项目摘要

9405803 Black Fatty acids are essential components of cellular membranes and are important sources of metabolic energy. This project focuses on the transport of long-chain fatty acids into cells followed by their enzymatic conversion to coenzyme A thioesters prior to metabolic utilization. In Escherichia coli, these two processes are postulated to be linked such that the esterification of long-chain fatty acids provides the driving force for transport. These compounds traverse the bacterial cell envelope by a high affinity, saturable, energy-dependent process that requires the outer membrane-bound fatty acid binding and transport protein FadL (product of the fadL gene) and the inner membrane associated acyl CoA synthetase (product of the fadD gene). Acyl CoA synthetase activates fatty acids concomitant with transport, thereby resulting in their net accumulation within the cell against a concentration gradient. The fadL and fadD genes have been cloned and sequenced and their respective gene products purified and characterized. In this work, the topology of FadL will be defined using partial proteolysis in isolated outer membranes and peptide purification and sequencing. Specific amino acid residues within FadL involved in fatty acid binding will be determined using chemical modification and site-directed mutagenesis. Amino acid residues within the fatty acid binding pocket within FadL will be modified using affinity labeling followed by proteolysis, peptide purification, and identification of the labeled amino acids. This will specifically define amino acids within FadL that comprise the fatty acid binding pocket and thus lay the groundwork for more detailed studies employing mutagenesis of the fadL gene. Collectively, these studies will define how FadL spans the outer membrane, and which specific amino acid residues sit within the fatty acid binding pocket. The role of acyl CoA synthetase in the transport of exogenous long-chain fatty acids will al so be further evaluated. Acyl CoA synthetase from E. coli has considerable similarity with its eukaryotic counterparts, particularly in a region of the enzyme postulated to bind ATP. Site-directed mutageneis of the acyl CoA synthetase structural gene will be used to define whether this region of the enzyme represents the ATP binding domain. %%% Fatty acids are the molecular building blocks of lipids. As such, they are essential components of cellular membranes, as well as important sources of metabolic energy. Depending on the cell type, the fatty acids needed by a cell may be either syntehsized by the cell itself, and/or taken up from the surrounding milieu. This project seeks to understand the biochemical mechanism whereby cells specifically take up fatty acids from their surroundings, using the common model bacterium, Escherichia coli, as a model system. A combination of genetic and biochemical approaches will be used. The results of these experiments are expected to expand our understanding of how fatty acids, and large molecules in general, are specifically transported across cell membranes. This kind of information is of fundamental importance to understanding how cells function. It is not difficult to see how this kind of information may have downstream application to a number of diverse areas of special interest; examples might include microbial physiological ecology, pharmacology, and animal or human nutrition. ***
小行星9405803 脂肪酸是细胞膜的重要组成部分,是代谢能量的重要来源。 该项目的重点是将长链脂肪酸运输到细胞中,然后在代谢利用之前将其酶促转化为辅酶A硫酯。 在大肠杆菌中,这两个过程被认为是相互关联的,使得长链脂肪酸的酯化提供了运输的驱动力。 这些化合物通过高亲和力、可饱和、能量依赖性过程穿过细菌细胞包膜,该过程需要外膜结合的脂肪酸结合和转运蛋白FadL(fadL基因的产物)和内膜相关的酰基CoA合成酶(fadD基因的产物)。 酰基辅酶A合成酶激活脂肪酸伴随着运输,从而导致它们在细胞内逆着浓度梯度的净积累。 fadL和fadD基因已被克隆和测序,其各自的基因产物纯化和表征。 在这项工作中,FadL的拓扑结构将被定义使用部分蛋白水解分离的外膜和肽纯化和测序。 将使用化学修饰和定点诱变来确定参与脂肪酸结合的FadL内的特定氨基酸残基。 FadL内脂肪酸结合口袋内的氨基酸残基将使用亲和标记进行修饰,然后进行蛋白水解、肽纯化和标记氨基酸的鉴定。 这将明确定义FadL内包含脂肪酸结合口袋的氨基酸,从而为采用fadL基因诱变的更详细研究奠定基础。 总的来说,这些研究将确定FadL如何跨越外膜,以及哪些特定的氨基酸残基位于脂肪酸结合口袋内。 酰基辅酶A合成酶在外源性长链脂肪酸转运中的作用也有待进一步研究。 E.大肠杆菌与其真核生物的对应物有相当大的相似性,特别是在假定与ATP结合的酶的区域。 酰基CoA合成酶结构基因的定点诱变将用于确定该酶区域是否代表ATP结合结构域。 %%%脂肪酸是脂质的分子构建块。 因此,它们是细胞膜的重要组成部分,也是代谢能量的重要来源。 根据细胞类型,细胞所需的脂肪酸可以由细胞自身合成,和/或从周围环境中摄取。 本项目以常见的模式细菌大肠杆菌为模型系统,研究细胞从周围环境中特异性摄取脂肪酸的生化机制。 将使用遗传和生物化学方法的组合。 这些实验的结果有望扩大我们对脂肪酸和一般大分子如何特异性跨细胞膜转运的理解。 这种信息对于理解细胞如何发挥功能至关重要。 不难看出这类信息如何在下游应用于许多不同的特别感兴趣的领域;例子可能包括微生物生理生态学,药理学和动物或人类营养学。 ***

项目成果

期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ monograph.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ sciAawards.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ conferencePapers.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ patent.updateTime }}

Paul Black其他文献

Progression in learning science
  • DOI:
    10.1007/bf02356878
  • 发表时间:
    1992-12-01
  • 期刊:
  • 影响因子:
    2.300
  • 作者:
    Paul Black;Shirley Simon
  • 通讯作者:
    Shirley Simon
Be Careful Who You Trust: Issues with the Public Key Infrastructure
小心你信任的人:公钥基础设施的问题
Immunotherapy of Metastatic Disease
转移性疾病的免疫治疗
Trabalhando por dentro da caixa preta: avaliação para a aprendizagem na sala de aula
Trabalhando por dentro da caixa preta: avaliação para a prendizagem na sala de aula
  • DOI:
    10.18676/cadernoscenpec.v8i2.445
  • 发表时间:
    2019
  • 期刊:
  • 影响因子:
    0
  • 作者:
    Paul Black;Christine Harrison;Clare Lee;Bethan Marshall;Dylan Wiliam
  • 通讯作者:
    Dylan Wiliam
Spam Email Categorization with NLP and Using Federated Deep Learning
通过 NLP 和联合深度学习进行垃圾邮件分类
  • DOI:
    10.1007/978-3-031-22137-8_2
  • 发表时间:
    2022
  • 期刊:
  • 影响因子:
    3.8
  • 作者:
    I. Haq;Paul Black;I. Gondal;J. Kamruzzaman;P. Watters;A. Kayes
  • 通讯作者:
    A. Kayes

Paul Black的其他文献

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

{{ truncateString('Paul Black', 18)}}的其他基金

Biochemistry of Fatty Acid Transport in Escherichia Coli
大肠杆菌脂肪酸运输的生物化学
  • 批准号:
    0331889
  • 财政年份:
    2003
  • 资助金额:
    $ 6.5万
  • 项目类别:
    Continuing grant
Biochemistry of Fatty Acid Transport in Escherichia Coli
大肠杆菌脂肪酸运输的生物化学
  • 批准号:
    0212745
  • 财政年份:
    2002
  • 资助金额:
    $ 6.5万
  • 项目类别:
    Continuing Grant
Biochemistry of FAtty Acid Transport In Escherichia Coli
大肠杆菌中脂肪酸运输的生物化学
  • 批准号:
    9816414
  • 财政年份:
    1999
  • 资助金额:
    $ 6.5万
  • 项目类别:
    Continuing Grant
Biochemistry of Fatty Acid Transport in Escherichia coli
大肠杆菌脂肪酸转运的生物化学
  • 批准号:
    9796006
  • 财政年份:
    1996
  • 资助金额:
    $ 6.5万
  • 项目类别:
    Continuing Grant
Biochemistry of Fatty Acid Transport in Escherichia coli
大肠杆菌脂肪酸转运的生物化学
  • 批准号:
    9506059
  • 财政年份:
    1995
  • 资助金额:
    $ 6.5万
  • 项目类别:
    Continuing Grant
Biochemistry of Fatty Acid Uptake in Escherichia coli
大肠杆菌摄取脂肪酸的生物化学
  • 批准号:
    9104646
  • 财政年份:
    1991
  • 资助金额:
    $ 6.5万
  • 项目类别:
    Continuing Grant
Biochemistry of Fatty Acid Uptake in Escherichia Coli
大肠杆菌摄取脂肪酸的生物化学
  • 批准号:
    8811714
  • 财政年份:
    1988
  • 资助金额:
    $ 6.5万
  • 项目类别:
    Continuing Grant
The Use of Geophysical Instruments in an Undergraduate Geology Curriculum
地球物理仪器在本科地质学课程中的使用
  • 批准号:
    7814958
  • 财政年份:
    1978
  • 资助金额:
    $ 6.5万
  • 项目类别:
    Standard Grant

相似国自然基金

FATTY ACID DESATURASE 4调节植物膜联蛋白活性的分子机制研究
  • 批准号:
    31870803
  • 批准年份:
    2018
  • 资助金额:
    55.0 万元
  • 项目类别:
    面上项目

相似海外基金

Biochemistry of fatty acid desaturases
脂肪酸去饱和酶的生物化学
  • 批准号:
    228861-2005
  • 财政年份:
    2009
  • 资助金额:
    $ 6.5万
  • 项目类别:
    Discovery Grants Program - Individual
Biochemistry of fatty acid desaturases
脂肪酸去饱和酶的生物化学
  • 批准号:
    228861-2005
  • 财政年份:
    2008
  • 资助金额:
    $ 6.5万
  • 项目类别:
    Discovery Grants Program - Individual
Biochemistry of fatty acid desaturases
脂肪酸去饱和酶的生物化学
  • 批准号:
    228861-2005
  • 财政年份:
    2007
  • 资助金额:
    $ 6.5万
  • 项目类别:
    Discovery Grants Program - Individual
Biochemistry of fatty acid desaturases
脂肪酸去饱和酶的生物化学
  • 批准号:
    228861-2005
  • 财政年份:
    2006
  • 资助金额:
    $ 6.5万
  • 项目类别:
    Discovery Grants Program - Individual
Biochemistry of fatty acid desaturases
脂肪酸去饱和酶的生物化学
  • 批准号:
    228861-2005
  • 财政年份:
    2005
  • 资助金额:
    $ 6.5万
  • 项目类别:
    Discovery Grants Program - Individual
Biochemistry of Fatty Acid Transport in Escherichia Coli
大肠杆菌脂肪酸运输的生物化学
  • 批准号:
    0331889
  • 财政年份:
    2003
  • 资助金额:
    $ 6.5万
  • 项目类别:
    Continuing grant
Biochemistry of fatty acid desaturases
脂肪酸去饱和酶的生物化学
  • 批准号:
    228861-2001
  • 财政年份:
    2003
  • 资助金额:
    $ 6.5万
  • 项目类别:
    Discovery Grants Program - Individual
Biochemistry of Fatty Acid Transport in Escherichia Coli
大肠杆菌脂肪酸运输的生物化学
  • 批准号:
    0212745
  • 财政年份:
    2002
  • 资助金额:
    $ 6.5万
  • 项目类别:
    Continuing Grant
Biochemistry of fatty acid desaturases
脂肪酸去饱和酶的生物化学
  • 批准号:
    228861-2001
  • 财政年份:
    2002
  • 资助金额:
    $ 6.5万
  • 项目类别:
    Discovery Grants Program - Individual
Biochemistry of fatty acid desaturases
脂肪酸去饱和酶的生物化学
  • 批准号:
    228861-2001
  • 财政年份:
    2001
  • 资助金额:
    $ 6.5万
  • 项目类别:
    Discovery Grants Program - Individual
{{ showInfoDetail.title }}

作者:{{ showInfoDetail.author }}

知道了