Unraveling the Reaction between Heme (FeIV=O) Moiety and H2S

揭示血红素 (FeIV=O) 部分与 H2S 之间的反应

• 批准号: 7284003
• 负责人: JUAN LOPEZ-GARRIGA
• 金额: $ 14.92万
• 依托单位: UNIVERSITY OF PUERTO RICO MAYAGUEZ
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7284003
• 关键词: Achievement; Affect; Amino Acid Sequence; Amino Acids; Bacteria; Bacterial Infections; Behavior; Binding; Binding Sites; Biological; Biophysics; Biopolymers; Biotechnology; Blood; Blood capillaries; Books; Caliber; Carbon Monoxide; Cations; Cell Aging; Cells; Characteristics; Chemicals; Chloride Ion; Chlorides; Clams; Complementary DNA; Complex; Condition; Coupled; Crystallization; Cyanides; DNA Sequence Rearrangement; Data; Data Collection; Databases; Dependence; Detection; Development; Diffusion; Discrimination; Disruption; Dissociation; Distal; Doctor of Philosophy; Education; Educational Curriculum; Educational process of instructing; Electronics; Engineering; Environment; Environmental Exposure; Erythrocytes; Escherichia coli; Frequencies; Globin; Glucose; Glutamine; Grant; Heme; Hemeproteins; Hemin; Hemoglobin; Hemoglobin J; Human; Hydrogen Bonding; Hydrogen Peroxide; Hydrogen Sulfide; Inorganic Chemistry; International; Investigation; Isopropyl Thiogalactoside; Journals; Kinetics; Knowledge; Laboratories; Lead; Length; Life; Ligand Binding; Ligands; Literature; Longitudinal Studies; Lucina pectinata hemoglobin I; Manuscripts; Measurement; Methodology; Methods; Modeling; Molecular; Molecular Conformation; Molecular Structure; Molecular Weight; Monitor; Movement; Mutate; Mutation; Myoglobin; Numbers; Object Attachment; Oxygen; Peripheral; Personal Satisfaction; Pliability; Population; Porphyrins; Positioning Attribute; Preparation; Procedures; Production; Property; Protein Chemistry; Proteins; Proteomics; Protons; Publications; Published Comment; Publishing; Puerto Rico; Pump; Pyrroles; Rate; Reaction; Recombinants; Reporting; Research; Research Project Grants; Resolution; Roentgen Rays; Role; Science; Score; Sepharose; Sequence Alignment; Sickle Cell Anemia; Site; Solutions; Solvents; Spectroscopy, Fourier Transform Infrared; Stream; Structure; Structure-Activity Relationship; Students; Sulfhemoglobin; Sulfides; Sulfur; System; Techniques; Thalassemia; Therapeutic; Time; Training; Tyrosine; Universities; Ursidae Family; Work; X ray spectroscopy; X-Ray Crystallography; absorption; abstracting; amino group; base; capillary; carbonyl group; cell age; chlorin; comparative; conformer; dimer; expression vector; flash photolysis; fluoromethyl 2,2-difluoro-1-(trifluoromethyl)vinyl ether; high school; improved; innovation; mutant; myoglobin cyanide; nanosecond; programs; protein expression; reaction rate; research study; sensor; size; stop flow technique; sulfheme; sulfmyoglobin; vibration

The peroxidative reactions of hemoglobin (Hb) and myoglobin (Mb) with hydrogen peroxide (H2O2) produce porphyrin /r-cation radical ferryl compound I (Fe'v=O Por*+) and ferryl compound II (Felv=O Por), which are very reactive and detrimental to red cells. At the same time, hydrogen sulfide (H2S) entering the blood stream, by environmental exposure or bacterial infections, can interact with these species leading to the formation of sulfhemoglobin (sulfHb) and sulfmyoglobin (sulfMb) with the subsequent disruption of Hb and Mb function. These inactive proteins, whose specific heme intermediates and reaction mechanism are still unknown, are characterized by a protoheme IX chlorin derivative with an absorption band at the 620 nm region, bearing the saturated 4-vinyl group with a H2S covalently bound across the¿-¿ double bond of the pyrrole "C". Our latest data show that under the same experimental conditions, of H202 and H2S, the hemoglobins I, II and III, (Hbl and Hbll/Hblll, respectively), from Lucina pectinata do not form these sulfheme derivatives. Hbl delivers H2S, while Hbll and Hblll transport 02 to symbiotic bacteria, despite the diversity in function and chemical structure no sulfheme-proteins derivatives have been detected in the L pectinata clam. This discrimination is not understood and will be used as a model to comprehend the mechanism toward sulfHb and sulfMb since the formation sulfHbl and sulfHbll/sulfHblll should proceed via the same high valent heme intermediates generated by the reaction with H2O2and H2S according to: H2S [Hbl (Fem) or Hbl (Fe"-O2) ] + H2O2~-* (Fe'v=O Por¿+) + (Felv=O Por) > SulfHbl (A) Compound I Compound II O2 Other notable difference between Hbl and Mb is that the ferryl compound I is near one thousand times more stable in the former than in the latter. Thus, our hypothesis suggests that the mechanism of sulfhemeprotein formation may depends on (1) the amino acid environment surrounding the heme center, (2) the life time of the ferryl species (compound I and II), and (3) the orientation and structure of the peripheral heme substituents. To explore these alternatives, we will monitor the bands at 648 nm, 620 nm, and 419 nm characteristic of Hbl compound I, sulfHbl, and compound II, respectively, by UV-Vis and stopped flow, upon the reaction (A) of hemeproteins (Mb, Hbl, Hbl mutants, and Hbll/Hblll). The intermediate and final structures will be pursued by resonance Raman, NMR and X-ray spectroscopy. The data will allow commenting on (i) the role of compound I and II in the formation of human sulfHb and sulfMb, (ii) unravel the selectivity of H2S for the pyrrole "C" reaction , and (iii) contribute to the development of direct strategies for the treatment and reversibility of sulfHb or sulfMb to the biological active human Hb and Mb.

血红蛋白 (Hb) 和肌红蛋白 (Mb) 与过氧化氢 (H2O2) 发生过氧化反应，产生 卟啉/r-阳离子自由基 Ferr1 化合物 I (Fe'v=O Por*+) 和 Ferr1 化合物 II (Felv=O Por)，它们是 对红细胞非常敏感且有害。与此同时，硫化氢（H2S）进入血液 通过环境暴露或细菌感染，水流可以与这些物种相互作用，导致 硫血红蛋白 (sulfHb) 和硫肌红蛋白 (sulfMb) 的形成，随后 Hb 和 MB 功能。这些无活性的蛋白质，其具体的血红素中间体和反应机制仍不清楚 未知，其特征在于原血红素 IX 二氢卟酚衍生物，吸收带位于 620 nm 区域，带有饱和的 4-乙烯基，其中 H2S 共价键合在 ¿-¿ 双键上 吡咯“C”。我们的最新数据表明，在相同的实验条件下，H202和H2S的 来自 Lucina pectinata 的血红蛋白 I、II 和 III（分别为 Hbl 和 Hbll/Hblll）不会形成这些 磺基衍生物。 Hbl 输送 H2S，而 Hbll 和 Hblll 将 02 输送给共生细菌，尽管 功能和化学结构的多样性 L 中未检测到硫血红素蛋白衍生物 梳状蛤。这种歧视不被理解，将被用作理解的模型 sulfHb 和 sulfMb 的机制，因为 sulfHbl 和 sulfHbll/sulfHblll 的形成应通过 根据以下公式，与 H2O2 和 H2S 反应生成相同的高价血红素中间体： 硫化氢 [Hbl (Fem) 或 Hbl (Fe"-O2) ] + H2O2~-* (Fe'v=O Por¿+) + (Felv=O Por) > SulfHbl (A) 化合物 I 化合物 II O2 Hbl 和 Mb 之间的其他显着区别是 ferryl 化合物 I 接近 1000 倍 前者比后者更稳定。因此，我们的假设表明 sulfheme 蛋白的机制 形成可能取决于（1）血红素中心周围的氨基酸环境，（2） Ferryl 物种（化合物 I 和 II）的寿命，以及 (3) 外周的方向和结构 血红素取代基。为了探索这些替代方案，我们将监测 648 nm、620 nm 和 419 nm 的波段 Hbl 化合物 I、sulfHbl 和化合物 II 分别通过 UV-Vis 和停止流动得到的 nm 特征， 血红素蛋白（Mb、Hbl、Hbl 突变体和 Hbll/Hblll）的反应 (A)。中间和最终 结构将通过共振拉曼、核磁共振和X射线光谱来追踪。数据将允许 评论 (i) 化合物 I 和 II 在人 sulfHb 和 sulfMb 形成中的作用，(ii) 阐明 H2S 对吡咯“C”反应的选择性，以及 (iii) 有助于直接策略的开发 用于治疗 sulfHb 或 sulfMb 并将其逆转为具有生物活性的人 Hb 和 Mb。