Outer sphere contributions within the thiol dioxygenase active site: a combined structure-function study

硫醇双加氧酶活性位点内外球的贡献：结构-功能组合研究

• 批准号: 10579496
• 负责人: Bradley S Pierce
• 金额: $ 41.37万
• 依托单位: UNIVERSITY OF ALABAMA IN TUSCALOOSA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-15 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10579496
• 关键词: Active Sites; Address; Affinity; Amines; Amino Acids; Anions; Binding; Binding Sites; Catalysis; Categories; Central Nervous System; Charge; Computing Methodologies; Conserved Sequence; Coupled; Cysteamine; Cysteine; Cysteine dioxygenase; Diagnostic; Dioxygen; Dioxygenases; Electron Transport; Embryo; Enzymes; Exhibits; GTP-Binding Protein Regulators; Health; Human; Hydrogen Bonding; Intuition; Investigation; Iron; Kinetics; Knowledge; Ligands; Maintenance; Malignant Neoplasms; Mammals; Measurement; Metabolic Diseases; Modeling; Molecular; Mononuclear; N-terminal; Neurodegenerative Disorders; Neurotransmitters; Osmoregulation; Oxygen; Oxygenases; Pathway interactions; Physiological; Play; Positioning Attribute; Proteins; Reaction; Reporting; Rheumatoid Arthritis; Role; Sampling; Scheme; Series; Site; Skeletal Muscle; Specificity; Structural Models; Structure; Substrate Specificity; Sulfhydryl Compounds; Sulfinic Acids; Taurine; bile salts; carboxylate; carboxylation; cross reactivity; cysteine sulfinic acid; cysteinyltyrosine; enzyme mechanism; enzyme substrate complex; experimental study; falls; heart rhythm; human disease; hypotaurine; iron nitrosyl; member; nervous system development; novel; oxidation; polypeptide; posttranscriptional; protein degradation; sensor; seryl-histidine; small molecule; spectroscopic survey; tool

TITLE. Outer sphere contributions within the thiol dioxygenase active site: a combined structure-function study PROJECT SUMMARY. Thiol dioxygenases (TDOs) are a subset of non-heme mononuclear iron oxygenases that catalyze the O2-dependent oxidation of thiol-bearing substrates to yield the corresponding sulfinic acid. Cysteine dioxygenase (CDO) and cysteamine dioxygenase (ADO) are the only known mammalian TDOs. These enzymes catalyze the oxidation of L-cysteine (CYS) and cysteamine (2-aminoethanthiol, CA) to produce cysteine sulfinic acid (CSA) and hypotaurine (HT), respectively. Ultimately, CSA and HT feed into the mammalian biosynthetic pathways for taurine (TAU). As the most abundant (sulfonic) amino acid derivative in the body, TAU plays a variety of essential functions including osmoregulation, conjugation substrate in the synthesis of bile salts, stabilization of skeletal muscle, maintenance of cardiac rhythm, and an essential neurotransmitter in embryonic central nervous system development. Despite having structurally similar substrates, CDO and ADO exhibit remarkable substrate-specificity; exhibiting no (or negligible) cross-reactivity. Significantly, changes in the cellular expression and/or activity of mammalian TDOs (CDO and ADO) have been correlated with the onset of various human diseases (cancer, neurodegenerative disorders, rheumatoid arthritis, as well as other metabolic disorders). An emerging subset of TDOs are referred to as N-terminal cysteinyl dioxygenases (NCOs). These enzymes are believed to function as physiologic O2-sensors by catalyzing the oxidation of protein N-terminal CYS-residues to initiate protein degradation. A similar function has recently been reported for mammalian ADO in controlling regulators of G protein signaling. Therefore ADO functions both as a small molecule TDO, specific for cysteamine (CA), as well as a NCO by regulating the stability of specific proteins in an O2-dependent fashion. A notable distinction within the NCO active site is the absence of a conserved sequence of spatially adjacent amino acids (Ser-His-Tyr) forming a hydrogen bonding network. This structural motif is highly conserved among structurally characterized TDOs and it is believed to regulate oxygen and thiol-substrate binding affinity at the TDO Fe-site. The absence of this conserved sequence among NCOs suggests a novel mechanism for O2-sensing in this class of enzymes. The central theme of the proposed activities is to investigate the role of outer-sphere interactions within the TDO active site that influence substrate efficiency and recognition. The experiments proposed involve a series of careful kinetic measurements to identify catalytically essential residues within the enzymatic active site. In parallel, spectroscopic and computational methods will be employed to develop a structural model for active site hydrogen-bonding interactions. Similar studies will be performed on members of the NCO class of enzymes to provide a valuable counterpoint to results obtained for TDOs. Potentially these studies will shed light on the mechanisms by which TDOs (and NCOs) regulate specificity of small molecule thiols, polypeptide substrates, and molecular oxygen.

标题。硫醇双加氧酶活性位点内的外球贡献：组合的结构功能 学习 项目摘要。硫醇双加氧酶 (TDO) 是非血红素单核铁加氧酶的一个子集 催化含硫醇底物的 O2 依赖性氧化，产生相应的亚磺酸。 半胱氨酸双加氧酶 (CDO) 和半胱胺双加氧酶 (ADO) 是唯一已知的哺乳动物 TDO。这些 酶催化 L-半胱氨酸（CYS）和半胱胺（2-氨基乙硫醇，CA）氧化产生半胱氨酸 分别是亚磺酸（CSA）和亚牛磺酸（HT）。最终，CSA 和 HT 进入哺乳动物体内 牛磺酸（TAU）的生物合成途径。作为体内最丰富的（磺酸）氨基酸衍生物， TAU 发挥多种重要功能，包括渗透调节、合成中的结合底物 胆汁盐、骨骼肌的稳定、心律的维持以及人体必需的神经递质 胚胎中枢神经系统发育。尽管 CDO 和 ADO 具有结构相似的底物 表现出显着的底物特异性；不表现出（或可忽略不计）交叉反应性。值得注意的是，变化 哺乳动物 TDO（CDO 和 ADO）的细胞表达和/或活性与发病相关 各种人类疾病（癌症、神经退行性疾病、类风湿性关节炎以及其他代谢性疾病） 障碍）。 TDO 的一个新兴子集被称为 N 末端半胱氨酰双加氧酶 (NCO)。这些 酶被认为通过催化蛋白质 N 末端的氧化而发挥生理 O2 传感器的作用 CYS 残基启动蛋白质降解。最近报道了哺乳动物 ADO 的类似功能 控制 G 蛋白信号传导的调节因子。因此，ADO 既具有小分子 TDO​​ 的功能，又具有特异性 半胱胺 (CA) 以及 NCO，通过以 O2 依赖性方式调节特定蛋白质的稳定性。 NCO 活性位点的一个显着区别是缺乏空间相邻的保守序列 氨基酸（Ser-His-Tyr）形成氢键网络。该结构基序在 具有结构特征的 TDO，据信可以调节氧和硫醇-底物的结合亲和力 TDO Fe-位点。 NCO 中这种保守序列的缺失表明了 O2 传感的新机制 在这一类酶中。拟议活动的中心主题是研究外层空间的作用 TDO 活性位点内的相互作用影响底物效率和识别。实验 建议涉及一系列仔细的动力学测量，以识别内的催化必需残基 酶活性位点。同时，将采用光谱和计算方法来开发 活性位点氢键相互作用的结构模型。类似的研究将对以下成员进行： NCO 类酶可为 TDO 获得的结果提供有价值的对照。潜在的这些 研究将揭示 TDO（和 NCO）调节小分子特异性的机制 硫醇、多肽底物和分子氧。

项目成果

Cooperative redox and spin activity from three redox congeners of sulfur-bridged iron nitrosyl and nickel dithiolene complexes.

• DOI: 10.1073/pnas.2201240119
• 发表时间: 2022-06-21
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1

Cyanide replaces substrate in obligate-ordered addition of nitric oxide to the non-heme mononuclear iron AvMDO active site.

• DOI: 10.1007/s00775-023-01990-7
• 发表时间: 2023-04
• 期刊: JOURNAL OF BIOLOGICAL INORGANIC CHEMISTRY
• 影响因子: 3
• 作者: York, Nicholas J. J.;Lockart, Molly M. M.;Schmittou, Allison N. N.;Pierce, Brad S. S.
• 通讯作者: Pierce, Brad S. S.

Outer-Sphere Tyrosine 159 within the 3-Mercaptopropionic Acid Dioxygenase S-H-Y Motif Gates Substrate-Coordination Denticity at the Non-Heme Iron Active Site.

• DOI: 10.1021/acs.biochem.9b00674
• 发表时间: 2019-12-24
• 期刊: BIOCHEMISTRY
• 影响因子: 2.9
• 作者: Sardar, Sinjinee;Weitz, Andrew;Hendrich, Michael P.;Pierce, Brad S.
• 通讯作者: Pierce, Brad S.

Nitroxyl Modified Tobacco Mosaic Virus as a Metal-Free High-Relaxivity MRI and EPR Active Superoxide Sensor.

• DOI: 10.1021/acs.molpharmaceut.8b00262
• 发表时间: 2018-08-06
• 期刊: Molecular pharmaceutics
• 影响因子: 4.9
• 作者: Dharmarwardana M;Martins AF;Chen Z;Palacios PM;Nowak CM;Welch RP;Li S;Luzuriaga MA;Bleris L;Pierce BS;Sherry AD;Gassensmith JJ
• 通讯作者: Gassensmith JJ

Low-Spin Cyanide Complexes of 3-Mercaptopropionic Acid Dioxygenase (MDO) Reveal the Impact of Outer-Sphere SHY-Motif Residues.

• DOI: 10.1021/acs.inorgchem.1c01519
• 发表时间: 2021-12-20
• 期刊: INORGANIC CHEMISTRY
• 影响因子: 4.6
• 作者: York, Nicholas J.;Lockart, Molly M.;Pierce, Brad S.
• 通讯作者: Pierce, Brad S.