Comparison between Myoglobin and Its Mutant(D60K) Interacting with Hydrogen Peroxide by Spectrum
ZHI Qiu-yan1,2, TANG Qian1, 2, CAO Hong-yu1, 2, AN Liang-mei2, ZHANG Ying-ying2, ZHENG Xue-fang1, 2*
1. Liaoning Key Lab of Bioorganic Chemistry, Dalian University, Dalian 116622, China 2. College of Bioengineering, Dalian University, Dalian 116622, China
Abstract:To characterize the roles played by surface-charged residue Asp60 in the structure stability of myoglobin when it was replaced with Lys, the interaction of myoglobin[Mb(WT)] and its mutant[Mb(D60K)] with hydrogen peroxide (H2O2) were studied by the method of ultraviolet-visible (UV-Vis) absorption spectroscopy, fluorescence spectroscopy and stopped-flow fluorescence spectroscopy under simulative physiological conditions. There are remarkable differences between Mb(D60K) and Mb(WT) in the UV-Vis absorption spectroscopy and fluorescence spectroscopy of iron porphyrin during the process of interaction. Although we only altered one external amino acide, the data showed that the function and structure stability of Mb(D60K) was greatly changed. Furthermore, results from synchronous fluorescence spectroscopy and stopped-flow fluorescence spectroscopy all indicated that H2O2 had less effect on the structure of Mb(D60K) while the structure of Mb(WT) was notably changed. From a comprehensive and comparative data analysis, the authors determined that the structure of Mb(D60K) was improved when it interacted with H2O2.
[1] Ozaki S, Roach M P, Matsui T, et al. Accounts of Chemical Research, 2001, 34: 818. [2] Raffaella R, Enrico M, Monica M. Biochemistry, 2004, 377: 717. [3] Lloyd E, Tu K M, Mauk A G, et al. American Chemical Society, 1995, 117: 6434. [4] Marshall S A, Morgan C S, Mayo S L. Jounral of Molecular Biology, 2002, 316: 189. [5] Egawa T, Yoshioka S, Takahashi S, et al. Biology Chemistry, 2003, 278: 41597. [6] Shiga T, Imaizumi K. Archives of Biochemistry and Biophysics, 1975, 167: 469. [7] Hoffman B M, Celis L M, Cull D A, et al. Proceedings of the National Academy of Sciences, 2005, 102: 3564. [8] LI Yi-wen, CAO Hong-yu, TANG Qian, et al(李宜雯,曹洪玉,唐 乾,等). Acta Physical-Chemistry(物理化学学报), 2010, 26(6): 1687. [9] Kanner J, Harel S. Archives of Biochemistry and Biophysics, 1985, 237: 314. [10] CHEN Guo-zhen,HUANG Xian-zhi,XU Jin-gou(陈国珍,黄贤智,许金钩). Principles of Fluorescence Scpetroscopy, Second Edition(荧光分析法, 第2版). Beinjing: Science Press(北京:科学出版社), 1990. 201. [11] Trynda-Lemiesz L, Karaczyn A, Keppler B K. Inorganic Biochemistry, 2000, 78(4): 341. [12] Zhu Changqing, Zhao Danhua, Chen Jinlong. Analatical and Bioanalalytical Chemistry, 2004, 378: 811. [13] MA Jun-yan, ZHENG Xue-fang, GUO Ming, et al(马君燕,郑学仿,郭 明, 等). Science in China Series B-Chemistry(中国科学, B辑-化学), 2008, 38(1): 55.