Mechanisms of Catalysis and Cofactor Biosynthesis of Redox Enzymes with Unusual Cofactors

具有异常辅因子的氧化还原酶的催化和辅因子生物合成机制

基本信息

批准号：
10544716
负责人：
VICTOR L DAVIDSON
金额：
$ 40.12万
依托单位：
UNIVERSITY OF CENTRAL FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-01-01 至 2023-07-31
项目状态：
已结题

项目摘要

This research program focuses on the characterization of redox enzymes with unusual cofactors, and their interactions with other electron transfer (ET) proteins. This includes enzymes that do not contain exogenous cofactors, but instead possess “protein-derived cofactors” that are formed by multiple irreversible post-translational modifications of amino acid residues. This research has made major contributions to at least three areas of broad biological and biomedical significance: elucidation of mechanisms of biological ET; characterization of mechanisms of biosynthesis of protein-derived cofactors and their catalytic properties; and discovery of novel mechanisms of heme function and oxygen activation. The future goals include characterization of three novel classes of redox enzymes that we have recently identified. Lod-A like proteins possess a cysteine tryptophylquinone (CTQ) cofactor derived from specific cysteine and tryptophan residues. These are unusual as they are the first enzymes with tryptophylquinone cofactors that function as oxidases rather than dehydrogenases. They are also the first amino acid oxidases described that uses a cofactor other than a flavin for catalysis. LodB-like proteins are flavoenzymes that catalyze the post-translational modifications required for CTQ biosynthesis on a precursor LodA-like protein. These reactions must be performed by “remote catalysis” that involves long-range ET, since the residues that are modified reside within the protein and are not surface exposed. As such, the mechanism of this process must be novel because no such reactions have been described that are catalyzed by a flavoenzyme, especially by remote catalysis. Rv2633c is a protein from Mycobacterium tuberculosis that is upregulated in response to the host defense during infection. It possesses two non-heme irons that are predicted from sequence to form an oxo-bridged hemerythrin-like site. We showed that this is the first hemerythrin-like protein to function as a catalase, and is the first example of a catalase with a non-heme di-iron cofactor. The results of these studies will further our understanding of the range and mechanisms of reactions that can be catalyzed by enzymes, particularly those involving oxygen reactivity and free radical intermediates. Understanding how enzymes control these reactive species during catalysis while minimizing oxidative damage will provide insights into how to mitigate the consequences of oxidative stress. This will also expand or current views about protein evolution and protein structure-function relationships, and provide insights for protein engineering strategies to introduce new functional groups into proteins.

该研究计划的重点是与不寻常的氧化还原酶的表征辅因子及其与其他电子转移（ET）蛋白的相互作用。这包括不包含外源辅助因子但具有“蛋白质来源的酶由多种不可逆的氨基酸翻译后修饰形成的辅助因子” 残留物。这项研究为至少三个广泛的生物学领域做出了重大贡献和生物医学意义：阐明生物ET机制；表征蛋白质衍生辅因子及其催化特性的生物合成的机制；和发现血红素功能和氧气激活的新型机制。未来目标包括我们最近拥有的三种新型氧化还原酶的表征确定。 LOD-A喜欢蛋白质具有半胱氨酸的色氨酸（CTQ）辅助因子。特异性半胱氨酸和色氨酸残留物。这些是不寻常的，因为它们是第一个酶色氨酸酮酮辅助因子作为氧化物而不是脱氢酶。他们是还描述了使用黄素以外的辅助因子进行催化的第一个氨基酸氧化物。 LODB样蛋白是黄酮酶，可催化所需的翻译后修饰用于前体洛氏蛋白上的CTQ生物合成。这些反应必须由涉及远程ET的“远程催化”，因为经过修改的残差位于蛋白质且不表面暴露。因此，这个过程的机制必须是新颖的因为尚未描述过由黄酮酶催化的这种反应，所以特别是通过远程催化。 RV2633C是结核分枝杆菌的一种蛋白质在感染期间对宿主防御的响应上调。它具有两个非血红素从序列预测的是形成氧桥的类似甲状腺肿的位点。我们表明了这一点这是第一个像过氧化氢蛋白酶作用的hegerythrin样蛋白过氧化氢酶，带有非血红素二铁辅因子。这些研究的结果将进一步了解可以被酶催化的反应的范围和机制，特别是涉及氧反应性和自由基中间体的人。了解如何酶在催化过程中控制这些反应性物种，而最小化氧化损伤将提供有关如何减轻氧化应激后果的见解。这也将扩大或有关蛋白质进化和蛋白质结构功能关系的当前观点，并提供蛋白质工程策略的见解，将新功能组引入蛋白质。

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Characterization of PlGoxB, a flavoprotein required for cysteine tryptophylquinone biosynthesis in glycine oxidase from Pseudoalteromonas luteoviolacea.

PlGoxB 的表征，PlGoxB 是黄黄假交替单胞菌甘氨酸氧化酶中半胱氨酸色氨酸醌生物合成所需的黄素蛋白。

DOI：
10.1016/j.abb.2019.108110
发表时间：
2019
期刊：
Archives of biochemistry and biophysics
影响因子：
3.9
作者：
Mamounis,KyleJ;Ma,Zhongxin;Sanchez-Amat,Antonio;Davidson,VictorL
通讯作者：
Davidson,VictorL

The hemerythrin-like diiron protein from Mycobacterium kansasii is a nitric oxide peroxidase.

DOI：
10.1016/j.jbc.2022.101696
发表时间：
2022-03
期刊：
The Journal of biological chemistry
影响因子：
0
作者：
Ma Z;Holland AA;Szlamkowicz I;Anagnostopoulos V;Caldas Nogueira ML;Caranto JD;Davidson VL
通讯作者：
Davidson VL

Structural Determinants of the Specific Activities of an L-Amino Acid Oxidase from Pseudoalteromonas luteoviolacea CPMOR-1 with Broad Substrate Specificity.

DOI：
10.3390/molecules27154726
发表时间：
2022-07-24
期刊：
MOLECULES
影响因子：
4.6
作者：
Mamounis, Kyle J.;Nogueira, Maria Luiza Caldas;Salvador, Daniela Priscila Marchi;Andreo-Vidal, Andres;Sanchez-Amat, Antonio;Davidson, Victor L.
通讯作者：
Davidson, Victor L.

Substitution of the sole tryptophan of the cupredoxin, amicyanin, with 5-hydroxytryptophan alters fluorescence properties and energy transfer to the type 1 copper site.

DOI：
10.1016/j.jinorgbio.2022.111895
发表时间：
2022-09
期刊：
JOURNAL OF INORGANIC BIOCHEMISTRY
影响因子：
3.9
作者：
Pastore, Anthony J.;Ficaretta, Elise;Chatterjee, Abhishek;Davidson, Victor L.
通讯作者：
Davidson, Victor L.

Diversity of structures and functions of oxo-bridged non-heme diiron proteins.