荧光光谱法测定二苯甲酮类卤酚与血红素加氧酶-1的相互作用  

Determination of Interaction between Benzophenone Halogenols and Heme Oxygenase-1 by Fluorescence Spectrometry

作　　者：刘金帅 宋婷[1] 冯秀娥[1,2] 李青山[1,2,3] LIU Jin-shuai;SONG Ting;FENG Xiu-e;Li Qing-shan(School of Pharmacy,Shanxi Medical University,Taiyuan 030001,China;Medicinal Basic Research Innovation Center of Chronic Kidney Disease,Ministry of Education,Shanxi Medical University,Taiyuan 030001,China;Shanxi Key Laboratory of Innovative Drug for the Treatment of Serious Diseases Basing on the Chronic Inflammation,Shanxi University of Chinese Medicine,Jinzhong 030619,China)

机构地区：[1]山西医科大学药学院,山西太原030001 [2]山西医科大学教育部慢性肾脏病医药基础研究创新中心,山西太原030001 [3]山西中医药大学基于炎性反应的重大疾病创新药物山西省重点实验室,山西晋中030619

出　　处：《化学研究与应用》2024年第7期1568-1576,共9页Chemical Research and Application

基　　金：重大新药创制国家科技重大专项项目(2018ZX09711001-001-017)资助;山西省重点研发计划项目(202102130501005)资助;山西省自然科学基金项目(201901D111209)资助;基于炎性反应的重大疾病创新药物山西省重点实验室2021年度开放课题项目(2021sxcxyw06)资助。

摘　　要：采用荧光光谱法,探讨三种具有强抗炎、抗氧化活性的二苯甲酮类卤酚3,4-二羟基-3'-氯二苯甲酮(LM46)、4,5-二羟基-2,3-二溴二苯甲酮(LM48)、2,4',5'-三羟基-5,2'-二溴二苯甲酮(LM49)与血红素加氧酶-1(HO-1)的相互作用机制,结合分子对接分析其相互作用模式。在模拟生理条件下,用荧光猝灭法求得卤酚与蛋白配合物的结合常数,探讨主要作用力类型,并用同步、三维荧光光谱探讨其对HO-1蛋白构象的影响。荧光猝灭实验结果表明,随着温度的升高,LM46、LM48、LM49的猝灭常数(K_(sv))均下降,表观结合常数(K_(b))相应降低。卤酚与HO-1的相互作用发生荧光猝灭的机制属于静态猝灭。热力学数据表明,LM46、LM48和LM49与HO-1间的作用主要为范德华力、氢键或质子化等作用力,且反应可自发进行。同步荧光光谱、三维荧光光谱结果表明,卤酚与HO-1结合后,使HO-1的构象发生变化,进而影响色氨酸(Trp)或酪氨酸(Tyr)残基周围的微环境,改变HO-1腔内的疏水环境。The interaction mechanism of three benzophenone halophenols 3,4-dihydroxy-3'-chlorobenzophenone(LM46),4,5-di-hydroxy-2,3-dibromobenzophenone(LM48),and 2,4',5'-trihydroxy-5,2'-dibromobenzophenone(LM49)possessing strongly anti-in-flammatory and antioxidant abilities with heme oxygenase-1(HO-1)protein was investigated by fluorescence spectroscopy,and their interaction mode was further analyzed by molecular docking.In simulated physiological conditions,the binding constants were ob-tained,and the interaction force was evaluated by fluorescence quenching method.Synchronous fluorescence and three-dimensional fluorescence spectra were further used to analyze the conformation changes of HO-1 during the interaction.The results of fluores-cence quenching experiment showed that the quenching constants(K_(sv))and the apparent binding constants(K_(b))were respectively decreased with the increase of temperature.The mechanism of fluorescence quenching in the process of interaction between halophe-nol and HO-1 was attributed to the static quenching.The thermodynamic data exhibited that the interaction force of compounds LM46,LM48,and LM49 with HO-1 was mainly Van der Waals force,hydrogen-bonding effect,and protonation.Moreover,the re-action could occur spontaneously.Synchronous and three-dimensional fluorescence spectra experiments presented that with the bind-ing of halophenols and HO-1,the conformation of HO-1 was changed to influence the micro-environment of tryptophan and tyrosine residues,and therefore changed the hydrophobic environment in HO-1 cavity.

关 键 词：荧光光谱 卤酚 血红素加氧酶-1 相互作用 分子对接 

分 类 号：O657.3[理学—分析化学]