|
|
|
|
|
|
| Sulfhydryl Modification and Spectral Measurement of Cytoglobin |
| ZHOU Dan-lei1, 2 |
1. School of Life Science, Beijing Institute of Technology, Beijing 100081, China
2. Davis Heart and Lung Research Institute, College of Medicine, The Ohio State University, Columbus, OH 43210, USA |
|
|
|
|
Abstract Cytoglobin (Cygb), a member of globin family, is a hemeprotein which was discovered 15 years ago. It contains a prosthetic group heme, which can reversibly bind an oxygen molecule between the iron ion of the porphyrin ring and a histidine of the polypeptide chain and play an important role in storing and delivering oxygen. There are two cysteines in C38 and C83 site of Cygb. Modification of the cysteine on Cygb can affect the oxygen-binding function of Cygb. In this article, chemical reagent 4, 4’- dithiodipyridine (4-PDS), N-ethylmaleimide (NEM), Oxidized Glutathione (GSSG) and Dithiothreitol (DTT) were used for modifying Cygb to intramolecular disulfide bond (Cygb-SS), thioether bonds (Cygb-SC), intermolecular disulfide bond (Cygb-SSG)and free sulfhydryls (Cygb-SH) respectively. The modified effect and product yield were detected by 4, 4’-dipyridine disulfide (4-PDS) spectroscopy method which can calculate modified rate by measuring the free sulfhydryl content in modified Cygb. The concentration of sulfhydryl in Cygb-SS and Cygb-SC samples is lower than one-tenth of the concentration of Cygb, indicating that the free sulfhydryl on the Cygb has been occupied by 4-PDS and NEM; The concentration of sulfhydryl in Cygb-SSG sample is comparable to that of the protein, indicating that one sulfhydryl in Cygb molecule participated in the reaction. Due to the steric hindrance of Cygb, the other sulfhydryl remained unchanged in the free sulfhydryl state; Double the concentration of sulfhydryl in the Cygb-SH sample as compared to the concentration of the Cygb indicates that one Cygb molecule contains two free sulfhydryl. We can know the modified yield by detecting the free sulfhydryl content of the four Cygbs respectively. The results showed that the four chemical reagents successfully modified cysteines on Cygb, and that the product yield reached more than 90%. This article measures the modified effect of Cygb by 4-PDS spectroscopy method and verifies the feasibility of this method by cysteine. In summary, the 4-PDS spectrometry method is accurate and reliable, which is complementary to the classical Ellman’s reagent method, and is more suitable for determining the content of sulfhydryl of compounds with absorption peak at 410~420 nm.
|
|
Received: 2018-02-24
Accepted: 2018-11-20
|
|
|
|
[1] Kawada N, Kristensen D B, Asahina K, et al. J. Biol. Chem., 2001, 276: 25318.
[2] Trent J T，Hargrove M S. J. Biol. Chem., 2002, 277: 19538.
[3] Burmester T, Ebner B, Weich B, et al. Mol. Biol. Evol., 2002, 19: 416.
[4] Tae B, Oliveira K C, Conceicao R R, et al. Environ. Toxicol., 2017, 32: 1252.
[5] Demirci S, Dogan A, Apdik H, et al. Mol. Cell. Biochem., 2018, 437: 133.
[6] Liu X P, El-Mahdy M A, Boslett J, et al. Nature Communications, 2017, doi: 10.1038/ncomms14807.
[7] Thuyle T T, Matsumoto Y, Thuy T T, et al. Am. J. Pathol., 2015, 185: 1045.
[8] Nishi H, Inagi R, Kawada N, et al. Am. J. Pathol., 2011, 178: 128.
[9] Liu X, Follmer D, Zweier J R, et al. Biochemistry，2012, 51：5072.
[10] Liu X, Tong J, Zweier J R, et al. FEBS J，2013, 280：3621.
[11] Van Thuy T T, Thuy L T, Yoshizato K, et al. Sci. Rep., 2017, 7: 41888.
[12] Thuyle T T, Van Thuy T T, Matsumoto Y, et al. Sci. Rep., 2016, 6: 24990.
[13] Wen J, Wu Y, Wei W, et al. Sci. Rep., 2017, 7: 41647.
[14] Oleksiewicz U, Liloglou T, Field J K, et al. Cell. Mol. Life Sci., 2011, 68: 3869.
[15] Reeder B J, Svistunenko D A, Wilson M T. The Biochemical Journal，2011, 434：483.
[16] ZHOU Dan-lei, LIU Xiao-ping, LUO Ai-qin(周丹蕾，刘小平，罗爱芹). SCIENTIA SINICA: Vitae(中国科学：生命科学)，2018，48(6)：630.
[17] Lechauve C, Chauvierre C, Dewilde S, et al. FEBS J，2010, 277：2696.
[18] Beckerson P, Reeder B J, Wilson M T. FEBS Lett., 2015, 589: 507.
[19] Tsujino H, Yamashita T, Nose A, et al. J. Inorg. Biochem., 2014, 135: 20.
[20] Zhou D, Hemann C, Boslett J, et al. FEBS Open Bio., 2017, 7: 845.
[21] Furuki K, Toyo’oka T. Anal. Biochem., 2017, 527: 33.
[22] Plancarte A, Nava G, Munguia J A. Exp. Parasitol., 2017, 177: 82.
[23] Corti P, Ieraci M, Tejero J. Nitric. Oxide., 2016, 53: 22.
[24] Rysman T, Jongberg S, Van Royen G, et al. J. Agric. Food Chem., 2014, 62: 12008.
[25] Liu X, Zweier J L. J. Electroanal. Chem. (Lausanne), 2013, 688: 32.
[26] Hamdane D, Kiger L, Dewilde S, et al. J. Biol. Chem., 2003, 278: 51713.
[27] Riener C K, Kada G, Gruber H. J.Anal. Bioanal. Chem., 2002, 373: 266.
[28] LIU Xin-rong, HONG Wei-wei, DENG Yi-hui(刘欣荣，洪维伟，邓意辉). Journal of Shenyang Pharmaceutical University(沈阳药科大学学报)，2013, 30：120. |
| [1] |
ZHENG Zhi-jie1, LIN Zhen-heng1, 2*, XIE Hai-he2, NIE Yong-zhong3. The Method of Terahertz Spectral Classification and Identification for Engineering Plastics Based on Convolutional Neural Network[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2023, 43(05): 1387-1393. |
| [2] |
ZHANG Zhong-xiong1, 2, 3, ZHANG Dong-li4, TIAN Shi-jie1, 2, 3, FANG Shi-yan1, 2, 3, ZHAO Yan-ru1, 2, 3*, ZHAO Juan1, 2, 3, HU Jin1, 2, 3*. Research Progress of Terahertz Spectroscopy Technique in Food Adulteration Detection[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2021, 41(05): 1379-1386. |
| [3] |
CHEN Yi-fan, LI Yun-jing, PENG Miao-miao, YANG Chun-yong*, HOU Jin, CHEN Shao-ping. Improvements of VIS-NIR Spectroscopy Model in the Prediction of TVB-N Using MIV Wavelength Selection[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2020, 40(05): 1413-1419. |
| [4] |
ZHANG Hu, HE Jian-ping*, LINYANG Sheng-lan. Three-Dimensional Arc Spectrum and Anti-Interference Decoupling in Micro Plasma Arc Welding[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2020, 40(01): 48-53. |
| [5] |
WANG Jin-hua1, CAO Lan-jie1, XU Guo-qiang2*, FENG Xiao-xin3, WU Bing1, ZHANG Bo1. Research on Hyper-Spectral Test of Concrete Corrosion Product under Sodium Sulfate Attack[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2019, 39(06): 1724-1730. |
| [6] |
LI Jing-hong1, 2, BIAN Lin2, TIAN Su-yan2, KONG Jin-ming3, LI Lian-zhi2* . Spectroscopic Study on the Interaction of Human Cytoglobin with Copper(Ⅱ) Ion[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2017, 37(01): 321-326. |
| [7] |
LI Jing1,2, LU Xu-tao1,2, Lü Hai-feng1,2, SUN Yun-qiang1,2 . Study on the Design of Explosive (HNS) Content Analysis System Based on THz Spectrum Monomer [J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2016, 36(11): 3506-3509. |
| [8] |
SONG Lin-li1, ZHOU Han-chang1, ZHANG Zhi-jie2. Research on a Novel High-Precision Methane Concentration Detection System[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2014, 34(12): 3354-3357. |
| [9] |
HU Yang-jun, ZHU Chun, CHEN Guo-qing*, ZHANG Yong, KONG Fan-biao, LI Run, ZHU Zhuo-wei, WANG Xu, GAO Shu-mei. Commercial Orange Juice Beverages Detection by Fluorescence Spectroscopy Combined with PCA-ED and PLSR Methods[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2014, 34(08): 2143-2147. |
| [10] |
WU Chen-kai, ZHANG Liang, SHEN Huang-tong, FU Xia-ping* . Investigation of Light Penetration Depth and Distribution Inside Citrus Tissue [J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2014, 34(03): 601-604. |
| [11] |
TAO Lin-li1, 2, YANG Xiu-juan1, 2, DENG Jun-ming1, 2, ZHANG Xi1, 2* . Application of Near Infrared Reflectance Spectroscopy to Predict Meat Chemical Compositions: A Review [J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2013, 33(11): 3002-3009. |
| [12] |
YAN Jie, MENG Peng-hua . Research on Methane Concentration Monitoring System Based on Electro-Optical Modulation Interference[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2013, 33(08): 2153-2156. |
| [13] |
CHEN Hui-hui, ZHU Duan-xu, GUO Yan-li*, WANG Ying, YAN Hong-tao . Study on Interaction of L-Homocysteine Modified Gold Nanoparticles with Bovine Serum Albumin by Fluorescence Spectroscopy [J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2012, 32(12): 3276-3280. |
| [14] |
CAO Hui1, HU Luo-na1, ZHOU Yan2* . Quantitative Analysis Method of Natural Gas Combustion Process Combining Wavelength Selection and Outlier Spectra Detection [J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2012, 32(10): 2799-2804. |
| [15] |
WANG Li-ying, MA Mei-hu*, HUANG Qun, SHI Xiao-xia. Study on the Stability of Chicken Egg Yolk Immunoglobulin (IgY) Modified with mPEG[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2012, 32(09): 2501-2507. |
|
|
|
|
