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New Nickel Environment for Biology: Cofactor Assembly and Function

生物学的新镍环境：辅因子组装和功能

基本信息

批准号：
1807073
负责人：
Robert Hausinger
金额：
$ 56.7万
依托单位：
Michigan State University
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2018
资助国家：
美国
起止时间：
2018-07-01 至 2022-06-30
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=1807073&HistoricalAwards=false
关键词：
New Nickel Environment Biology Cofactor

项目摘要

With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Dr. Robert Hausinger and Dr. Jian Hu from Michigan State University to characterize how a microorganism interconverts the two mirror image structures of lactic acid, a central element of carbohydrate metabolism. The enzyme that does this, called lactate racemase, requires both a protein component and nickel. Nickel is very rarely found in biology. Even more remarkably, the nickel in lactate racemase is present in a unique environment never before observed. This project investigates how the bacterium makes the cofactor that creates the unique environment. This effort provides training to a postdoctoral researcher, a graduate student, and undergraduate students in the areas of structure and biochemical mechanisms. These researchers also participate in hands-on educational activities for the public related to this topic that will engage lifelong learners of all ages at the annual Science Festival at the university.This research elucidates structure-function aspects of the four proteins required for lactate racemase activity. It characterizes how LarB catalyzes a new reaction, carboxylation of the pyridinium ring of nicotinic acid adenine dinucleotide while hydrolyzing the phosphoanhydride linkage to produce pyridinium-3,5-dicarboxylic acid mononucleotide. Additional studies of LarE, a sacrificial sulfur transferase, establish how the two carboxylic acid groups are transformed into thiocarboxylate groups and define whether the resulting dehydroalanine side chain of the enzyme can be regenerated to cysteine. This work uncovers how LarC catalyzes the CTP-dependent installation of Ni into pyridinium-3-5-dithiocarboxylic acid mononucleotide, the first biological example of a cyclometallation reaction, to create the cofactor. Finally, these investigations show how the cofactor is bound to different LarA species and identifies other potential transformations catalyzed by the enzyme. In combination, the biosynthesis and roles of this novel nickel-containing cofactor can be described. This innovative research expands our understanding of nickel biochemistry and establishes a new paradigm for redox chemistry involving a tethered nickel-pincer cofactor.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

有了这个奖项，化学部的生命过程化学项目资助密歇根州立大学的Robert Hausinger博士和Jian Hu博士研究微生物如何相互转化乳酸的两种镜像结构，乳酸是碳水化合物代谢的核心元素。这样做的酶，称为乳酸消旋酶，需要蛋白质成分和镍。镍在生物学中非常罕见。更值得注意的是，乳酸消旋酶中的镍存在于一个以前从未观察到的独特环境中。该项目研究了细菌如何制造创造独特环境的辅因子。这项工作提供了培训，以博士后研究人员，研究生和本科生在结构和生化机制的领域。这些研究人员还参加了与此主题相关的公众实践教育活动，这些活动将在大学的年度科学节上吸引所有年龄段的终身学习者。这项研究阐明了乳酸消旋酶活性所需的四种蛋白质的结构-功能方面。它表征了LarB如何催化一个新的反应，烟酸腺嘌呤二核苷酸的吡啶环的羧化，同时水解磷酸酐键产生吡啶-3，5-二羧酸单核苷酸。对LarE（一种牺牲性硫转移酶）的其他研究确定了两个羧酸基团如何转化为硫代羧酸基团，并确定了酶的脱氢丙氨酸侧链是否可以再生为半胱氨酸。这项工作揭示了LarC如何催化CTP依赖性的安装镍到吡啶-3-5-二硫代羧酸单核苷酸，第一个生物的例子，环金属化反应，以创建辅因子。最后，这些调查显示，辅因子是如何绑定到不同的LarA物种，并确定其他潜在的转化催化的酶。结合，这种新的含镍辅因子的生物合成和作用可以描述。这项创新性的研究扩展了我们对镍生物化学的理解，并为涉及拴系镍钳辅因子的氧化还原化学建立了新的范例。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

期刊论文数量（11）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Structural and mutational characterization of a malate racemase from the LarA superfamily

LarA 超家族苹果酸消旋酶的结构和突变特征

DOI：
10.1007/s10534-022-00372-x
发表时间：
2022
期刊：
BioMetals
影响因子：
3.5
作者：
Gatreddi, Santhosh;Urdiain-Arraiza, Julian;Desguin, Benoît;Hausinger, Robert P.;Hu, Jian
通讯作者：
Hu, Jian

Lanthanide-dependent alcohol dehydrogenases require an essential aspartate residue for metal coordination and enzymatic function

DOI：
10.1074/jbc.ra120.013227
发表时间：
2020-06-12
期刊：
JOURNAL OF BIOLOGICAL CHEMISTRY
影响因子：
4.8
作者：
Good, Nathan M.;Fellner, Matthias;Martinez-Gomez, N. Cecilia
通讯作者：
Martinez-Gomez, N. Cecilia

Analysis of the Active Site Cysteine Residue of the Sacrificial Sulfur Insertase LarE from Lactobacillus plantarum

DOI：
10.1021/acs.biochem.8b00601
发表时间：
2018-09-25
期刊：
BIOCHEMISTRY
影响因子：
2.9
作者：
Fellner, Matthias;Rankin, Joel A.;Hausinger, Robert P.
通讯作者：
Hausinger, Robert P.

The LarB carboxylase/hydrolase forms a transient cysteinyl-pyridine intermediate during nickel-pincer nucleotide cofactor biosynthesis

DOI：
10.1073/pnas.2106202118
发表时间：
2021-09-28
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
影响因子：
11.1
作者：
Rankin, Joel A.;Chatterjee, Shramana;Hausinger, Robert P.
通讯作者：
Hausinger, Robert P.

Crystallographic characterization of a tri-Asp metal-binding site at the three-fold symmetry axis of LarE