| 光谱学与光谱分析 |
|
|
|
|
|
| Research on the Reaction Dynamics between Peroxynitrite and Tyrosine Catalyzed with Mimic Enzyme through Flow Injection Analyzer |
| AN Xue-jing, LUO Yun-jing*, ZHANG Chen-yang |
| College of Life Science and Bioengineering, Beijing University of Technology, Beijing 100124, China |
|
|
|
|
Abstract As a highly active free radical in vivo, peroxynitrite can damage various biological macromolecules and cause a series of major diseases, which is of great significance to determine its content and reactive mechanism. It is very difficult to capture the dynamic process of peroxynitrite due to its active property and fast reacted rate. In this paper, we firstly explored the kinetic characteristics of peroxynitrite and tyrosine with the presence of Hemoglobin and Hemin by using flow injection analyzer. The results showed that the oxidation processes of peroxynitrite and tyrosine catalyzed with hemoglobin and hemin were in accordance with Michaelis-Menten’s dynamics law; Based on the Michaelis constant (Km) and the maximum initial rate (Vmax), we deduced the reaction mechanism that peroxynitrite, catalyzed by mimic enzymes, directly oxidized tyrosine combined with mimic enzymes, rather than decomposed to ·OH and O-2·; In addition, we detected the rate constant of the reaction catalyzed by these two kinds of enzymes at different temperature and pH, resulting that the optimum conditions of hemoglobin to catalyze this system were 25 ℃ and pH 8, the rate constant was 1.035×106 mol·L-1·s-1, while hemin was 37 ℃ and pH 9.5, the rate constant was 6.842×105 mol·L-1·s-1; Comparing the kinetic parameters, KmHb(4.46 μmol·L-1)<KHeminm(4.90 μmol·L-1), VHbmax(0.072 ΔIF/s)>VHeminmax(0.026 ΔIF/s), we discovered that the rate constant of hemoglobin in optimum condition was greater than that in hemin, and the catalytic activity of hemoglobin was higher than that of hemin. All these results have provided kinetic parameters for the study of determining the content of peroxynitrite and exploring its reaction mechanism with enzymatic method. It also laid a theoretical foundation for developing new diagnosis technology to prevent and cure diseases caused by free radicals in vivo.
|
|
Received: 2015-12-14
Accepted: 2016-04-19
|
|
|
|
Corresponding Authors:
LUO Yun-jing
E-mail: luoyj@bjut.edu.cn
|
|
[1] Szhbo C, Ischiropoulos H, Radi R. Nature Reviews Drug Discovery, 2007, 6(8): 662.
[2] WANG Rui-yong, KANG Xiao-hui, WU Jing, et al(王瑞勇，康小慧，吴 静，等). J. Zhengzhou Univ. (Nat. Sci. Ed.) (郑州大学学报(理学版)), 2012, 44(4): 78.
[3] Sarah C R, Lorenzo B. Proc. SPIE, 2013, 8579(10): 13.
[4] Shu Y L, Ming M D, Li Z, et al. Sensors and Actuators, 2016, 22: 198.
[5] Yun Z, Jing L F, Jin F N, et al. Biosensors and Bioelectronics, 2015, 73: 13.
[6] Yue S, Hong D, Yu T L, et al. Biosensors and Bioelectronics, 2016, 77: 894.
[7] Sara R, Israel P, Mordechai S, et al. Angewandte Chemie, 2015, 54(42): 12379.
[8] YAN Shuang, LI Xiao-yan, LIU Zhi-chang, et al(颜 双，李晓燕，刘志昌，等). Physical Testing and Chemical Analysis(Part B: Chemical Analysis)(理化检测(化学分册)), 2015, 51(3): 300.
[9] LIU Shan-du, YANG Xue-wei, ZHOU Wu-xiong, et al(刘珊渡，杨学蔚，周武雄，等). Applied Chemical Induatry(应用化工), 2014, 43(3): 552.
[10] Huang Qiuli, Yang Qianqian, Chen Yahong, et al. Henan Chemical Industry, 2015, 23(2): 55.
[11] Michael K, Jorgen A, Christin G, et al. Nature Communications, 2015, 6(7644): 1.
[12] LI Yong-sheng, ZHENG Wei, GAO Xiu-feng(李永生，郑 魏，高秀峰). Chinese Journal of Analytical Chemistry(分析化学), 2015, 43(10): 1580. |
| [1] |
GAO Fei1, XIONG Yang2 , ZHANG Ming1, ZHU Shao-ling3 . Investigations on NIR-SERS Spectra of Oxyhemoglobin for Lung Cancer Based on NIR-SERS Substrate [J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2017, 37(02): 441-445. |
| [2] |
LUO Man1*, HUANG Yuan-yuan1, SU Bao-chang2, SHI Yun-feng3, ZHANG Hong1, YE Xun-da1 . Study on RBC Oxygen-Carrying Function with the Incubation Time [J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2015, 35(12): 3350-3355. |
| [3] |
WANG Wei, PAN Zhi-feng*, TANG Wei-yue, LI Yun-tao, FAN Chun-zhen . Research on Early Diagnosis of Gastric Cancer by the Surface Enhanced Raman Spectroscopy of Human Hemoglobin [J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2015, 35(12): 3402-3405. |
| [4] |
LI Yan-qin1, MU Lan1, ZENG Xi1*, LI Jun1, Takehiko Yamato2 . The Thiacalix[4]arene-Coumarin Fluorescence Probe Recognition for Fe3+ and Bovine Hemoglobin [J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2015, 35(11): 3111-3116. |
| [5] |
XU Gang1, LI Xiao-li1*, LIU Xiao-min2 . A Simple Design of Functional Near-Infrared Spectroscopy System [J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2015, 35(02): 552-556. |
| [6] |
LIN Ling1, LI Wei1, ZHOU Mei1, ZENG Rui-li2, LI Gang3, ZHANG Bao-ju4* . Application of EMD Algorithm to the Dynamic Spectrum Non-Invasive Measurement of Hemoglobin[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2014, 34(08): 2106-2111. |
| [7] |
ZHAO Jing1, LIN Ling2, LU Xiao-zuo1, LI Gang2* . Noninvasive Detection of Hematocrit and the Mean Corpuscular Hemoglobin Concentration Levels by Vis-NIR Spectroscopy[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2014, 34(03): 652-655. |
| [8] |
LU Ming-zi1, GUO Yan-jun2, ZHAO Lian1*, ZHOU Hong1* . Application of Raman Spectroscopy to the Research on Hemoglobin Structure and Function[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2014, 34(02): 439-444. |
| [9] |
CAO Jun-tao1, WANG Hui1, CHEN Yong-hong2, LIU Yan-ming1* . Sensitive Determination of Hemoglobin in Human Blood and Serum by Flow Injection Coupled with Chemiluminescence Detection [J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2014, 34(01): 241-245. |
| [10] |
ZHOU Xue1, CHEN Guang-yi2, ZHANG Jian-min1, FAN Chun-zhen1, LIANG Er-jun1, GUO Yan2, TANG Wei-yue1* . Research on Early Diagnosis of Esophageal Cancer by Raman Spectroscopy of Human Hemoglobin[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2013, 33(11): 2989-2992. |
| [11] |
GUO Shi-jun1, ZENG Chang-chun1, 2*, LI Li-jun2, NIE Guang3, LIU Song-hao4 . Quantization Determination Study of Micro-Raman Spectroscopy of Methemoglobin Induced by Sodium Nitrite[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2013, 33(07): 1805-1809. |
| [12] |
LIU Pei-pei1, GUAN Xiao-yue1, ZENG Chang-chun1, 2*, NIE Guang3, LIU Song-hao4 . Raman Spectroscopy Study of the Effect of H+ on the Oxygen Affinity Capacity of Hemoglobin[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2013, 33(05): 1234-1238. |
| [13] |
FAN Yi-chen1, 2, LU Qi-peng1*, DING Hai-quan1, GAO Hong-zhi1, CHEN Xing-dan1 . Study on the Application of Empirical Mode Decomposition to Noninvasive Hemoglobin Measurement by NIRS[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2013, 33(02): 349-353. |
| [14] |
XIONG Yang1, LI Yun-tao1, GUO Yan2, RAO Feng-fei1, ZHANG Jian-min1, SI Min-zhen3, LIU Ren-ming3, TANG Wei-yue1*. Analysis of Surface Enhanced Raman Scattering Spectra of Oxyhemoglobin for Liver Cancer with Combined Multivariate Statistics[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2012, 32(09): 2427-2432. |
| [15] |
LI Jin-jing1, TANG Qian1,2, CAO Hong-yu1,2, ZHANG Yu-jiao1, ZHANG Tao1, ZHENG Xue-fang1,2* . Spectroscopic Studies of Guanidine Hydrochloride-Induced Unfolding of Hemoglobin[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2012, 32(09): 2496-2500. |
|
|
|
|
