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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

基本信息

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
项目类别：
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

项目摘要

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：结构-功能结合

项目成果

期刊论文数量（16）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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
作者：
通讯作者：

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.