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Interaction Between Cytochrome c and the Hapten 2,4-Dinitro-fluorobenzene by Electrospray Ionization Mass Spectrometry
Bo Wu,Yan-qiu Chu,Zhao-yun Dai,Chuan-fan Ding¤
Author NameAffiliationE-mail
Bo Wu Physical Chemistry Institute, Department of Chemistry, Fudan University, Shanghai 200433, China  
Yan-qiu Chu Physical Chemistry Institute, Department of Chemistry, Fudan University, Shanghai 200433, China  
Zhao-yun Dai Infectious Disease Department, Huadong Hospital, Shanghai 200040, China  
Chuan-fan Ding¤ Physical Chemistry Institute, Department of Chemistry, Fudan University, Shanghai 200433, China cfding@fudan.edu.cn, 
Abstract:
Allergic contact dermatitis is a delayed hypersensitivity reaction, which results from skin exposure to low molecular weight chemicals such as haptens. To clarify the pathogenic mechanism, electrospray ionization mass spectrometry (ESI-MS) and hydrogen/deuterium (H/D) exchange, as well as UV spectroscopy, were applied to determine the interaction between the model protein cytochrome c (cyt c) and the hapten 2,4-dinitro-fluorobenzene (DNFB). The ESI-MS results demonstrate that the conformation of cyt c can change from native folded state into partially unfolded state with the increase of DNFB. The equilibrium state H/D exchange followed by ESI-MS further confirms the above results. UV spectroscopy indicates that the strongfield coordination between iron of heme (prosthetic group) and His18 or Met80 of cyt c is not obviously affected by the hapten.
Key words:  Interaction, Cytochrome c, 2,4-dinitro-fluorobenzene, ESI-MS
FundProject:国家自然科学基金
Interaction Between Cytochrome c and the Hapten 2,4-Dinitro-fluorobenzene by Electrospray Ionization Mass Spectrometry
吴波,储艳秋,戴兆云,丁传凡*
摘要:
采用电喷雾质谱、H/D交换和紫外光谱测定了半抗原2,4-二硝基氟苯和模型蛋白细胞色素c的相互作用. 电喷雾质谱结果表明,随着2,4-二硝基氟苯体积比百分含量的增加,细胞色素c的构象可以由天然的折叠构象转变为部分非折叠构象,最后转变为完全非折叠构象.细胞色素c在平衡态的H/D交换结合电喷雾质谱实验结果进一步确认了上述构象变化. 紫外光谱结果表明半抗原的加入对辅基血红素上的亚铁离子和细胞色素c上His18和Met80的强场配位作用没有明显的影响.
关键词:  相互作用,细胞色素c,2,4-二硝基氟苯,电喷雾质谱
DOI:10.1088/1674-0068/21/03/217-220
分类号: