摘要
α-葡萄糖苷酶(GAA)作为一类重要的糖苷水解酶,是维持人体血糖平衡的重要功能性蛋白质。长期以来,为实现高血糖人群的降糖,研究人员多关注于探寻具有GAA活性抑制作用的食品、药品,但鲜有研究关注于人体非主动摄取的外源物质对GAA正常生理功能可能产生的影响。该研究选取典型多环芳烃类物质—菲的羟基代谢产物9-羟基菲(9-OHPhe),探究其对GAA活性功能的影响及潜在机制。为获得9-OHPhe与GAA的结合作用信息,PARAFAC法被应用于光谱重叠的9-OHPhe和GAA的三维荧光光谱(EEM)数据的解析,同时分子对接法被运用于分析分子层面两者结合的微观信息。9-OHPhe对GAA的活性有抑制作用,相应的IC50值为(7.59±1.91)μmol L^(-1);PARAFAC可有效应用于荧光光谱重叠的GAA和9-OHPhe反应体系的荧光数据解析;9-OHPhe可引起GAA内源荧光的静态猝灭,两者能形成1∶1复合物,在298 K下其结合常数为(8.91±0.68)×10^(3) L·mol^(-1);9-OHPhe与GAA的GLN281、LEU305、ASN319、LEU321、TYR322、LEU323和TYR352之间产生了疏水作用力,与GLN309和ASN319分别形成了键长为2.71和3.05Å的氢键;低浓度的9-OHPhe可使GAA的二级结构变得稳定,但过高浓度的9-OHPhe则会破坏GAA的结构稳定性。9-OHPhe对GAA活性有明显抑制效应,提示相关外源污染物进入健康人体后经这一途径所可能引起的血糖平衡紊乱值得关注和深入研究。
Alpha-glucosidase(GAA),as an important glycoside hydrolase,is avital functional protein to maintain human blood glucose balance.For a long time,researchers have focused on exploring foods and drugs with inhibition ofα-glucosidase activity in order to reduce blood glucose in people with hyperglycemia.However,few studies have focused on the possible effects of non-active intake of exogenous substances on the normal physiological functions of GAA.Given this,9-hydroxyphenanthrene(9-OHPhe),a hydroxyl metabolite of a typical polycyclic aromatic hydrocarbon—phenanthrene,was selected in this study to explore its influence on GAA activity and potential mechanism.In order to obtain the binding information of 9-OHPhe and GAA,the PARAFAC method was applied to analyze the three-dimensional fluorescence spectra(EEM)data of 9-OHPhe and GAA with overlapping spectra,and the molecular docking method was used to analyze the microscopic information of their binding at the molecular level.9-OHPhe inhibited the activity of GAA,and the corresponding IC50 value was(7.59±1.91)μmol·L^(-1).PARAFAC method can be effectively used to analyze the fluorescence data of GAA and 9-OHPhe systems with overlapping fluorescence spectra.9-OHPhe can cause the static quenching of endogenous fluorescence of GAA,and they can form a 1∶1 complex with the binding constant of(8.91±0.68)×10^(3) L·mol^(-1) at 298 K.The hydrophobic interactions existed between 9-OHPhe and GLN281,LEU305,ASN319,LEU321,TYR322,LEU323,TYR352 in GAA,and two hydrogen bonds with bond lengths of 2.71 and 3.05Åformed for 9-OHPhe with GLN309 and ASN319.A low concentration of 9-OHPhe can stabilize the secondary structure of GAA,while too high a concentration of 9-OHPhe will destroy the structural stability of GAA.9-OHPhe showed anobvious inhibitory effect on the activity of GAA,suggesting that the glucose balance disorder which might be caused through this pathway after the relevant exogenous pollutants entered the healthy human body was worthy of attention and in-depth study.
作者
张静
王鸿辉
金亮
廖颖敏
李恒
ZHANG Jing;WANG Hong-hui;JIN Liang;LIAO Ying-min;LI Heng(Key Laboratory of Estuarine Ecological Security and Environmental Health,School of Environmental Science and Engineering,Xiamen University Tan Kah Kee College,Zhangzhou 363105,China)
出处
《光谱学与光谱分析》
SCIE
EI
CAS
CSCD
北大核心
2024年第2期398-405,共8页
Spectroscopy and Spectral Analysis
基金
国家自然科学基金项目(22108231)
福建省自然科学基金项目(2021J01044)
漳州市自然科学基金项目(ZZ2020J19)资助。
关键词
Α-葡萄糖苷酶
9-羟基菲
三维荧光光谱
平行因子分析法
结合作用
活性抑制
α-Glucosidase
9-Hydroxyphenanthrene
Three-dimensional fluorescence spectroscopy
Parallel factor analysis
Binding interaction
Activity inhibition
