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血清蛋白对麦角甾-4,6,8,22-四烯-3-酮荧光增强作用的研究及应用
  • ISSN号:0567-7351
  • 期刊名称:《化学学报》
  • 时间:0
  • 分类:O622.4[理学—有机化学;理学—化学]
  • 作者机构:[1]西北大学化工学院,西安710069, [2]西北大学生命科学院,西安710069, [3]西北大学物理系,西安710069
  • 相关基金:国家自然科学基金(No.81001622)、科技部“重大新药创制”科技专项(No.2011ZX09401-308-34)资助项目.
中文摘要:

麦角甾-4,6,8,22.四烯-3-酮(ergone)为猪苓中主要甾体之一,具有多种药理和生理活性.研究了不同极性溶剂中ergone的光学性质,考察了人血清蛋白(HsA),牛血清蛋白(BSA)对ergone荧光和紫外光谱的影响,结果表明血清蛋白加入后ergone的光谱信号强度显著增强,并发生蓝移.根据Benesi-Hildebrand方程求得了结合常数和自由能变.基于血清蛋白对ergone具有良好的荧光增强作用,在模拟生理条件下以ergone为荧光探针,建立了一种灵敏的蛋白质定量分析方法,HSA和BSA线性响应浓度范围分别为(0.38~16.67)×10^-mol·L-1和(0.42~15.25)×10^-7mol·L-1,检测限(3们分别为1.01×10^-10和1.22×10^-10mol·L-1.考察共存物质对测定结果的影响中发现Fe抖会显著淬灭ergone荧光.该方法用于人血清中总蛋白含量测定结果与考马斯亮蓝法基本一致.

英文摘要:

Ergosta-4,6,8(14),22-tetraen-3-one (ergone) from many medicinal plants has been demonstrated to possess a variety of pharmacological activities in vivo and in vitro, including cytotoxic, diuretic and im- munosuppressive activity. The effect of different solvents on spectral characteristic of ergone was investi- gated. The interactions between ergone and human serum albumin (HSA) and bovine serum albumin (BSA) had been studied by using absorption and fluorescence spectroscopy. Absorption and fluorescence spectral studies showed that binding to the serum albumins leaded to a blue shift of ergone together with a notable intensity change. Furthermore, the number of binding sites (n) was identified by the absorption spectra. The binding constant (Ka) and the free energy changes (AG) were obtained by analysis of fluorescence data of the ergone in the absence and presence of HSA and BSA according to Benesi-Hildebrand equation. Compared to BSA, HSA associated with ergone in a stronger way. A new fluorescence quantitative determination of proteins has been developed by using ergone as a fluorescence probe. Good calibration curves of the proteins were obtained in the range of (0.38~6.67)×10^-7 mol,L-1 for HSA with detection limits (3tr) of 1.01 × 10-10 mol·L-1, and (0.42~15.25)×10^-7 moloL-1 for BSA with detection limits of 1.22×10-10 mol·L-1. Most metal ions had no notable effect on the determination of proteins except Fe3+ ions which could quench the fluorescence intensity of ergone. Determination of proteins in human serum by this method gave results which were very close to those obtained by Coomassie Brilliant Blue colorimetry.

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期刊信息
  • 《化学学报》
  • 北大核心期刊(2014版)
  • 主管单位:中国科学院
  • 主办单位:中国化学会 中国科学院上海有机化学研究所
  • 主编:周其林
  • 地址:上海市零陵路345号
  • 邮编:200032
  • 邮箱:hxxb@sioc.ac.cn
  • 电话:021-54925085
  • 国际标准刊号:ISSN:0567-7351
  • 国内统一刊号:ISSN:31-1320/O6
  • 邮发代号:4-209
  • 获奖情况:
  • 首届国家期刊奖,第二届国家期刊奖提名奖,中国期刊方阵“双高期刊”
  • 国内外数据库收录:
  • 俄罗斯文摘杂志,美国化学文摘(网络版),荷兰文摘与引文数据库,美国科学引文索引(扩展库),日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),英国英国皇家化学学会文摘,中国北大核心期刊(2000版)
  • 被引量:28694