Spectral Study on the Interaction between Cytochrome c and ProliNONOate
TANG Qian1, 2, GONG Ting-ting1, 2, SHI Shan-shan1, 2, CAO Hong-yu1, 2, WANG Li-hao3, YU Yong3, SHAN Ya-ming3, ZHENG Xue-fang1, 2*
1. School of Life Science and Biotechnology, Dalian University, Dalian 116622, China
2. Liaoning Key Lab of Bio-organic Chemistry, Dalian 116622, China
3. College of Life Sciences, Jilin University, Changchun 130012, China
Abstract：It played an important role in the detection of mitochondrial apoptosis that the interaction of Cytochrome(Cyt c) with NO, thus the research is still a hotspot issue for the chemists and biologists. In this paper, Ultraviolet (UV-Vis) absorbance spectra, UV-Vis time course spectra, circular dichroism (CD), synchronous fluorescence spectra and electron paramagnetic resonance (EPR) technology were used to research the coordination reaction mechanism of different valence states Cyt c with proliNO/NO and its spatial conformation changes. The results showed that Cyt c could directly react with NO without other reagents. And its secondary structure would change during the experiment. It has been established that the electronic configurations of iron ions in porphyrin complexes are controlled in the nature by a number of axial ligands, peripheral substituents attached to the porphyrin macrocycle, deformation of the porphyrin ring and solvent effects. During the experiment, proliNO/NO was added to the sample of Cyt c, and NO gas would generate, then entered into the solution, finally the distal methionine heme ligand was displaced, Cyt c could bind NO, that is, the Fe—S broke down, and the Fe-N formed. At the same time, generating a new cytochrome c-NO (Cyt c-NO) complex. Cyt c-NO binary complexes were instability and would dissociate, and dissociation process of Cyt c-NO binary complexes was belonging to a first-order reaction with the dissociation rate of (0.071 1±0.039 6) s-1. The secondary structure of Cyt c was affected by proliNO/NO concentration. When the concentrate of proliNO/NO was below to 8.6×10-4 mol·L-1, the peak changes of 222 nm and 208 nm was very weak, and the α-helix increased from 33.1% to 44.1%. And it continued to increase, the secondary structure of Cyt c took place a great change. The tiny changes illustrated that a new compound generated, but excessive proliNO/NO can break the structure of Cyt c. Taken together, we have demonstrated that an understanding of the interaction mechanism of NO with cytc, and the reaction mechanism of NO with ferri- and ferro-cytochrome c have important implications for the inhibition of mitochondrial oxygen consumption by NO and the mitochondrial metabolism of NO.
唐 乾，宫婷婷，史珊珊，曹洪玉，王立皓，于 勇，单亚明，郑学仿. 光谱法研究细胞色素C与proliNONOate之间的相互作用[J]. 光谱学与光谱分析, 2017, 37(08): 2505-2512.
TANG Qian, GONG Ting-ting, SHI Shan-shan, CAO Hong-yu, WANG Li-hao, YU Yong, SHAN Ya-ming, ZHENG Xue-fang. Spectral Study on the Interaction between Cytochrome c and ProliNONOate. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2017, 37(08): 2505-2512.
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