食品科学

• 基础研究 • 上一篇    下一篇

一磷酸腺苷与肌动球蛋白相互作用的荧光光谱研究

张 淼1,2,王道营1,*,张牧焓1,杨玉玲2,卞 欢1,吴海虹1,诸永志1,耿志明1,徐为民1,3   

  1. 1.江苏省农业科学院农产品加工研究所,江苏 南京 210014;2.南京财经大学食品科学与工程学院,江苏 南京 210046;3.江苏省肉类生产与加工质量安全控制协同创新中心,江苏 南京 210095
  • 出版日期:2016-09-15 发布日期:2016-09-22

Fluorescence Spectroscopic Study of the Molecular Interaction between Adenosine 5’-Monophosphate and Actomyosin

ZHANG Miao1,2, WANG Daoying1,*, ZHANG Muhan1, YANG Yuling2, BIAN Huan1, WU Haihong1, ZHU Yongzhi1, GENG Zhiming1, XU Weimin1,3   

  1. 1. Institute of Agricultural Products Processing, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China;
    2. College of Food Science and Engineering, Nanjing University of Finance and Economics, Nanjing 210046, China;
    3. Jiangsu Collaborative Innovation Center of Meat Production and Processing, Quality and Safety Control, Nanjing 210095, China
  • Online:2016-09-15 Published:2016-09-22

摘要:

目的:采用荧光分光光度法研究了一磷酸腺苷(adenosine 5’-monophosphate,AMP)与肌动球蛋白相互作用的情况。方法:根据分子间的相互作用力与相关热力学参数间的关系,结合AMP与肌动球蛋白相互作用的焓变和熵变,讨论AMP对肌动球蛋白荧光的猝灭机理,测定AMP与肌动球蛋白的猝灭常数、结合常数和作用力类型,考察AMP对肌动球蛋白构象的影响。结果:Western blotting证明了不同浓度AMP对肌动球蛋白解离的促进作用;AMP与肌动球蛋白结合反应中ΔG<0,ΔH>0,ΔS>0,两者之间结合主要通过疏水相互作用,且AMP的存在使得肌动球蛋白的构象发生变化,疏水环境极性降低;荧光光谱分析表明AMP与肌动球蛋白的结合使肌动球蛋白内源荧光发生了有规律的猝灭,298 K和313 K温度条件下AMP与肌动球蛋白的静态猝灭常数分别为8.70×102、4.07×102 L/mol,结合常数KLB分别为9.253×102 L/mol和3.142×102 L/mol。结论:AMP通过疏水相互作用力和肌动球蛋白形成复合物,从而促进肌动球蛋白的解离。

关键词: 肌动球蛋白, 一磷酸腺苷, 荧光猝灭, 解离

Abstract:

Objective: To study the interaction between adenosine 5’-monophosphate (AMP) and actomyosin by
fluorophotometry. Methods: Based on the correlation between intermolecular interaction and related thermodynamic
parameters, as well as enthalpy change (ΔH) and entropy change (ΔS) for their interaction, the fluorescence quenching
mechanism of actomyosin by AMP was discussed. Quenching constant, binding equilibrium constant and the number of
binding sites were determined. The effect of AMP on the conformation of actomysin was also analyzed using synchronous
fluorescence spectrometry. Results: Western blotting demonstrated the promotion effect of different concentrations of AMP on
the dissociation of actomyosin; the combination between them were mainly via hydrophobic interaction, since ΔG < 0, ΔH > 0,
ΔS > 0 were found in the binding reaction of AMP and actomyosin, and the presence of AMP changes the conformation of
actomyosin, reduce the polar of hydrophobic environment; Fluorescence spectroscopy showed that the combination of AMP and
actomyosin makes the regular quenching of intrinsic fluorescence of actomyosin. At 298 K and 313 K, the KSV of combination
were 8.70 × 102 and 4.07 × 102 L/mol, KLB were 9.253 × 102 L/mol and 3.142 × 102 L/mol respectively. Conclusion: Different
concentrations of AMP can promote the dissociation of actinmyosin as demonstrated by Western blotting.

Key words: actomyosin, adenosine 5’-monophosphate, fluoresence quenching, dissociation

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