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Novel Catalases and their Products

新型过氧化氢酶及其产品

基本信息

批准号：
8450914
负责人：
ALAN R. BRASH
金额：
$ 31.67万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-04-01 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8450914
关键词：
Active Sites Address Aerobic Amino Acids Anabaena Anabolism Area Atherosclerosis Binding Biochemistry Biology Carbon Catalysis Catalytic Domain Characteristics Chemistry Cyanobacterium Cyclization Cytochrome P450 Cytochromes DNA Sequence Rearrangement Diabetes Mellitus Dioxygenases Disease Enzyme Interaction Enzymes Epoxy Compounds Exhibits Family member Fatty Acids Felis catus Fusarium Grant Heme Hemeproteins Hydrogen Peroxide Hydrolysis Hydroxylation Inflammation Knowledge Life Ligands Light Lipid Peroxides Malignant Neoplasms Metabolic Metabolism Mixed Function Oxygenases Nature Nomenclature Nostoc Organism Oxidants Oxidative Stress Oxides Oxygen Peroxides Property Protein Family Proteins Pseudomonas fluorescens Reaction Relative (related person)Role Sentinel Sequence Homology Site-Directed Mutagenesis Specificity Structure Synthesis Chemistry System Thinking Tyrosine Work allene oxide synthase catalase cyclooxygenase 2 cyclopentenone enzyme activity heme a improved insight interest novel polypeptide propadiene public health relevance

项目摘要

DESCRIPTION (provided by applicant): The long-term objective of this application is to elucidate the roles of a novel group of proteins related in sequence and structure to catalase and to define the biochemistry and chemistry of their reactions and novel products. Catalase is renowned for its efficient reaction with hydrogen peroxide and its key role in the oxidative defense of all aerobic organisms. The catalase-related relatives studied here are smaller proteins with reaction specificity directly against fatty acid hydroperoxides. The prototypical enzyme of the proposal is an allene oxide synthase, an enzyme that catalyzes a cytochrome P450-type of reaction yet which exhibits distinct sequence homology to catalase. In Aim 1, the ability to transform the catalase-related AOS and the P450 type of AOS to monooxygenases will be examined using surrogate oxygen donors to activate the heme, with stopped flow spectral recording to detect short-lived intermediates as well as detailed product analysis to define the substrate-enzyme interaction. We also propose to change the AOS activity of these enzymes by site-direct mutagenesis of active site amino acids, a hypothesis supported by the array of products of CYP74 relatives of the P450 AOS. Aim 2 will focus on analysis of the catalytic activities of other novel enzyme candidates that have similar sequence characteristics defined as retention of the heme-binding features of a catalase within an unusually short polypeptide for catalases of only ~40kD. Aim 3 will analyze the mechanism of biosynthesis of a unique bicyclobutane fatty acid made by a catalase-related enzyme from the cyanobacterium Anabaena, and also characterize the structures of its hydrolysis and rearrangement products. This has implications regarding the potential synthesis of bicyclobutanes in other areas of biology. We will also address structural issues pertinent to the cyclization of natural allene oxide diastereomers, which is of fundamental interest in understanding the nature of allene oxide metabolism and the chemistry of cyclopentenone synthesis. The results of this study will provide new insights and a new way of thinking about the enzymatic capabilities of a long-recognized protein family with established roles as a sentinel at the forefront of oxidative defense.

描述（由申请人提供）：本申请的长期目标是阐明在序列和结构上与过氧化氢酶相关的一组新蛋白质的作用，并定义其反应和新产物的生物化学和化学。过氧化氢酶以其与过氧化氢的有效反应而闻名，并在所有有氧生物的氧化防御中发挥关键作用。这里研究的过氧化氢酶相关亲缘是较小的蛋白质，具有直接针对脂肪酸氢过氧化物的反应特异性。该提议的原型酶是一种氧化丙烯合成酶，这种酶催化细胞色素p450型反应，但与过氧化氢酶具有明显的序列同源性。在Aim 1中，将使用替代供氧体来激活血红素，以检测将过氧化氢酶相关的AOS和P450型AOS转化为单加氧酶的能力，并使用停止流光谱记录来检测短寿命中间体以及详细的产物分析来定义底物-酶相互作用。我们还提出通过位点直接诱变活性位点氨基酸来改变这些酶的AOS活性，这一假设得到了P450 AOS的一系列CYP74亲缘产物的支持。Aim 2将重点分析其他具有相似序列特征的新型候选酶的催化活性，这些特征被定义为过氧化氢酶在异常短的过氧化氢酶多肽中保留血红素结合特征，仅为~40kD。目的3将分析一种独特的双环丁烷脂肪酸的生物合成机制，该双环丁烷脂肪酸是由一种来自水蓝藻的过氧化氢酶相关酶合成的，并表征其水解和重排产物的结构。这对其他生物学领域中潜在的双环丁烷合成具有启示意义。我们还将讨论与天然氧化亚烯非对映体的环化有关的结构问题，这对理解氧化亚烯代谢的性质和环戊酮合成的化学性质具有重要意义。这项研究的结果将提供新的见解和新的方式来思考一个长期公认的蛋白质家族的酶促能力，并在氧化防御的前沿确立了哨兵的角色。

项目成果

期刊论文数量（13）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

A 49-kDa mini-lipoxygenase from Anabaena sp. PCC 7120 retains catalytically complete functionality.

来自鱼腥藻属的 49 kDa 小型脂氧合酶。

DOI：
10.1074/jbc.m705780200
发表时间：
2008
期刊：
The Journal of biological chemistry
影响因子：
0
作者：
Zheng,Yuxiang;Boeglin,WilliamE;Schneider,Claus;Brash,AlanR
通讯作者：
Brash,AlanR

Mechanistic aspects of CYP74 allene oxide synthases and related cytochrome P450 enzymes.

DOI：
10.1016/j.phytochem.2009.08.005
发表时间：
2009-09
期刊：
Phytochemistry
影响因子：
3.8
作者：
Brash AR
通讯作者：
Brash AR

DOI：
10.1016/bs.mie.2018.02.019
发表时间：
2018
期刊：
Methods in enzymology
影响因子：
0
作者：
Brash AR
通讯作者：
Brash AR

Omega-3 fatty acids are oxygenated at the n-7 carbon by the lipoxygenase domain of a fusion protein in the cyanobacterium Acaryochloris marina.

DOI：
10.1016/j.bbalip.2009.09.004
发表时间：
2010-01
期刊：
Biochimica et biophysica acta
影响因子：
0
作者：
Gao B;Boeglin WE;Brash AR
通讯作者：
Brash AR

珊瑚中与过氧化氢酶相关的血红素蛋白专门用于合成短链醛：过氧化氢酶中 P450 型氢过氧化物裂解酶活性的发现。

DOI：
10.1074/jbc.m115.660282
发表时间：
2015
期刊：
The Journal of biological chemistry
影响因子：
0
作者：
Teder,Tarvi;Lõhelaid,Helike;Boeglin,WilliamE;Calcutt,WadeM;Brash,AlanR;Samel,Nigulas
通讯作者：
Samel,Nigulas