| 游咏,刘芳,高淑琴,林英武,文格波.D-核糖诱导人肌红蛋白快速非酶糖基化.[J].中南医学科学杂志.,2016,(5):499-503, 535. |
| D-核糖诱导人肌红蛋白快速非酶糖基化 |
| D-ribose Induced Rapid Non-enzymatic Glycation of Human Myoglobin |
| 投稿时间：2016-03-26 修订日期：2016-05-31 |
| DOI： |
| 中文关键词: 核糖 葡萄糖 肌红蛋白 非酶糖基化 |
| 英文关键词:D-ribose D-glucose Myoglobin non enzymatic glycation |
| 基金项目:湖南省教育厅资助科研项目(13C817),湖南省卫生计生委科研计划课题项目(C2016041). |
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| 中文摘要: |
| 目的 探讨D-核糖与D-葡萄糖对人肌红蛋白(HMb)分子构象的影响,并研究反应后晚期糖基化终末产物(AGEs)的形成情况,以期为糖尿病及其并发症的防治提供新的研究线索。方法表达、纯化获得HMb,在体外分别与D-核糖及D-葡萄糖共孵育,采用紫外可见光谱(UV-vis)监测反应后HMb的结构变化,利用荧光光谱法监测反应后AGEs的生成情况。通过SDS-PAGE观察HMb与D-核糖及D-葡萄糖反应后分子量的变化情况。结果UV-vis显示D-核糖可在短时间内导致HMb血红素活性中心构象发生改变,且呈时间依赖性；同时随着核糖糖化时间的延长,具自发荧光的AGEs生成也逐渐增多。SDS-PAGE结果显示D-核糖能很快地与HMb发生反应,形成高分子量多聚体。而在相同的反应条件下,D-葡萄糖对HMb构象的影响及AGEs的形成显著较D-核糖为弱。结论D-核糖,而不是D-葡萄糖,能更快速地诱导HMb发生非酶糖基化,形成AGEs。 |
| 英文摘要: |
| Objective To investigate the effect of D-ribose and D-glucose on molecular conformation of human myoglobin(HMb),determine the formation of advanced glycation end products (AGEs) through in vitro glycation,and provide new clues for the prevention and treatment of diabetes mellitus and its complications.MethodHMb was expressed in BL21(DE3),purified,and incubated with D-ribose or D-glucose.Structural changes of HMb were studied by using UV-Vis spectroscopy.The formation of AGEs was monitored by Fluorescence spectrum.The molecular weight changes of HMb after reaction were observed by SDS-PAGE.ResultsIt showed that the heme active site of HMb was altered by interactions with D-ribose in a time-dependent manner.Simultaneously,the specific fluorescence of AGEs was found to increase markedly with increasing the incubation time of HMb incubated with D-ribose.SDS-PAGE showed that HMb rapidly reacted with D-ribose and formed higher aggregates.However,D-glucose has slight effects on the structural change of HMb and the formation of AGEs under the same conditions.ConclusionIn summary,this study shows that D-ribose,but not D-glucose,rapidly induces ribosylation of human myoglobin and generates AGEs. |
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