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: 7816959
• 负责人: JUAN LOPEZ-GARRIGA
• 金额: $ 12.2万
• 依托单位: UNIVERSITY OF PUERTO RICO MAYAGUEZ
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7816959
• 关键词: Affect; Amino Acid Sequence; Amino Acids; Bacteria; Bacterial Infections; Behavior; Binding; Binding Sites; Biological; Biophysics; Biotechnology; Blood; Blood capillaries; Books; Caliber; Carbon Monoxide; Cations; Cell Aging; Cells; Characteristics; Chemicals; Chloride Ion; Chlorides; Clams; Complementary DNA; Complex; Coupled; Crystallization; Cyanides; DNA Sequence Rearrangement; Data; Data Collection; Databases; Dependence; Detection; Development; Diffusion; Discrimination; Dissociation; Distal; Doctor of Philosophy; Education; Educational Curriculum; Educational process of instructing; Electronics; Engineering; Environment; Environmental Exposure; Erythrocytes; Escherichia coli; Frequencies; Globin; Glucose; Glutamine; Heme; Hemeproteins; Hemin; Hemoglobin; Hemoglobin J; Human; Hydrogen Bonding; Hydrogen Peroxide; Hydrogen Sulfide; Inorganic Chemistry; International; Isopropyl Thiogalactoside; Journals; Kinetics; Knowledge; Laboratories; Lead; Length; Life; Ligand Binding; Ligands; Longitudinal Studies; Lucina pectinata hemoglobin I; Manuscripts; Measurement; Methodology; Methods; Modeling; Molecular; Molecular Conformation; Molecular Structure; Molecular Weight; Monitor; Movement; Mutate; Mutation; Myoglobin; Oxygen; Peripheral; Population; Porphyrins; Positioning Attribute; Preparation; Production; Property; Protein Chemistry; Proteins; Proteomics; Protons; Publications; Published Comment; Publishing; Puerto Rico; Pump; Pyrroles; Reaction; Recombinants; Reporting; Research; Research Project Grants; Resolution; Roentgen Rays; Role; Science; Sepharose; Sequence Alignment; Sickle Cell Anemia; Site; Solutions; Solvents; Spectroscopy, Fourier Transform Infrared; Stream; Structure; Structure-Activity Relationship; Students; Sulfhemoglobin; Sulfides; System; Techniques; Thalassemia; Therapeutic; Time; Training; Tyrosine; Universities; Ursidae Family; Work; X ray spectroscopy; X-Ray Crystallography; absorption; abstracting; amino group; base; capillary; cell age; chlorin; conformer; dimer; expression vector; flash photolysis; flexibility; fluoromethyl 2,2-difluoro-1-(trifluoromethyl)vinyl ether; high school; improved; innovation; meetings; mutant; myoglobin cyanide; nanosecond; pi bond; programs; protein expression; reaction rate; research study; sensor; sulfheme; sulfmyoglobin; treatment strategy

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）的过氧化反应产生 卟啉 /r-cation自由基轮渡化合物I（Fe'V = O Por*+）和Ferryl化合物II（FELV = O POR），是 非常反应性，对红细胞有害。同时，硫化氢（H2S）进入血液 通过环境暴露或细菌感染，流可以与这些物种相互作用 磺胺血红蛋白（硫B）和磺基莫洛蛋白（硫基）的形成，随后Hb和Hb的破坏 MB功能。这些非活性蛋白，其特异性血红素中间体和反应机制仍然是 未知的特征是原蛋白IX氯素衍生物，在620 nm处具有吸收带 区域，具有饱和的4-乙烯基，与H2S共价结合了跨该区域 吡咯“ C”。我们的最新数据表明，在相同的实验条件下，H202和H2S， 血红蛋白I，II和III（分别来自Lucina Pectinata）不形成这些 亚硫衍生物。 HBL提供H2S，而HBLL和HBLL运输02到共生细菌，dospite 功能和化学结构的多样性未在L中检测到硫蛋白衍生物 果肉蛤。这种歧视尚不理解，将被用作理解的模型 由于形成硫和硫酸/硫酸盐/硫磺的机制应通过 根据：与H2O2和H2S产生的相同的高节质血红素中间体： H2S [hbl（fem）或hbl（fe“ -o2）] + h2o2〜-*（fe'v = oporòpor？ +） +（felv = o por）> sulfhbl（a） 化合物I化合物II O2 HBL和MB之间的另一个值得注意的区别在于，Ferryl Implound I接近一千次 前者比后者更稳定。这就是我们的假设表明硫蛋白质的机制 形成可能取决于（1）血红素中心周围的氨基酸环境，（2） 渡轮物种的寿命（化合物I和II），以及（3）外围的方向和结构 血红素子标准。为了探索这些替代方案，我们将在648 nm，620 nm和419中监视乐队 HBL化合物I，Sulfhbl和化合物II的NM特征分别由UV-VIS和停止流动， 在反应（a）时，血管蛋白（MB，HBL，HBL突变体和HBLL/HBLLLL）。中级和最终 共振拉曼，NMR和X射线光谱法将追求结构。数据将允许 评论（i）化合物I和II在人类硫和硫的形成中的作用，（ii）解开 H2S对吡咯“ C”反应的选择性，以及（iii）有助于发展直接策略 为了治疗和可逆性，硫B或硫对生物活性人体HB和MB的治疗和可逆性。