牛α-乳白蛋白-亞油酸復(fù)合物的結(jié)構(gòu)及抗腫瘤活性
發(fā)布時間:2018-12-14 00:44
【摘要】:利用內(nèi)源性熒光光譜、熒光探針(ANS)結(jié)合熒光光譜及圓二色譜,以乳白蛋白-油酸為參照,研究了乳白蛋白結(jié)合亞油酸后,疏水性氨基酸、疏水性區(qū)域、三級結(jié)構(gòu)及二級結(jié)構(gòu)的變化,并利用亞甲基藍的方法評價了該復(fù)合物的抗腫瘤活性。熒光光譜結(jié)果顯示,與乳白蛋白-油酸復(fù)合物類似,乳白蛋白結(jié)合亞油酸后,其內(nèi)源性熒光光譜顯著紅移,從331.07nm移至337.60nm;外源性ANS結(jié)合光譜藍移,從516.20nm移至508.50nm;且熒光強度增加,表明乳白蛋白結(jié)合亞油酸后同樣出現(xiàn)了疏水性氨基酸及疏水性區(qū)域暴露的現(xiàn)象。圓二色譜結(jié)果表明,與乳白蛋白-油酸復(fù)合物類似,乳白蛋白結(jié)合亞油酸后,三級結(jié)構(gòu)部分喪失,二級結(jié)構(gòu)中β-轉(zhuǎn)角及無規(guī)卷曲的含量顯著降低,β-折疊含量增加。細胞實驗證實了乳白蛋白-亞油酸復(fù)合物具有良好的抗腫瘤效果。該研究從復(fù)合物結(jié)構(gòu)和功能的兩方面,為新型抗腫瘤乳白蛋白-亞油酸復(fù)合物的開發(fā)提供了依據(jù)。
[Abstract]:The hydrophobic amino acids and hydrophobic regions of lactalbumin binding linoleic acid were studied by using endogenous fluorescence spectrum, fluorescence probe (ANS) binding fluorescence spectrum and circular dichroism, with lactoalbumin oleic acid as reference. The antitumor activity of the complex was evaluated by methylene blue method. The fluorescence spectra showed that, similar to the lactoalbumin-oleic acid complex, the endogenous fluorescence spectra of lactalbumin bound linoleic acid shifted significantly from 331.07nm to 337.60 nm. The blue shift of exogenous ANS binding spectrum from 516.20nm to 508.50 nm, and the increase of fluorescence intensity, indicated that hydrophobic amino acids and hydrophobic region exposures also appeared after the binding of lactalbumin to linoleic acid. The results of circular dichroism showed that, similar to lactoalbumin-oleic acid complex, the tertiary structure was partially lost after lactalbumin binding to linoleic acid, the 尾 -rotation angle and the content of random coil in the secondary structure decreased significantly, and the content of 尾 -fold increased. Cell experiments confirmed that lactoalbumin-linoleic acid complex had a good anti-tumor effect. This study provides a basis for the development of a novel anti-tumor lactoalbumin-linoleic acid complex in terms of its structure and function.
【作者單位】: 北京工商大學食品學院;國家糧食局科學研究院;中國農(nóng)業(yè)大學教育部-北京市共建功能乳品重點實驗室;
【基金】:國家科技支撐計劃項目(2012BAD12B08) 國家國際科技合作專項(2011DFA32550)資助
【分類號】:R914;R96
本文編號:2377568
[Abstract]:The hydrophobic amino acids and hydrophobic regions of lactalbumin binding linoleic acid were studied by using endogenous fluorescence spectrum, fluorescence probe (ANS) binding fluorescence spectrum and circular dichroism, with lactoalbumin oleic acid as reference. The antitumor activity of the complex was evaluated by methylene blue method. The fluorescence spectra showed that, similar to the lactoalbumin-oleic acid complex, the endogenous fluorescence spectra of lactalbumin bound linoleic acid shifted significantly from 331.07nm to 337.60 nm. The blue shift of exogenous ANS binding spectrum from 516.20nm to 508.50 nm, and the increase of fluorescence intensity, indicated that hydrophobic amino acids and hydrophobic region exposures also appeared after the binding of lactalbumin to linoleic acid. The results of circular dichroism showed that, similar to lactoalbumin-oleic acid complex, the tertiary structure was partially lost after lactalbumin binding to linoleic acid, the 尾 -rotation angle and the content of random coil in the secondary structure decreased significantly, and the content of 尾 -fold increased. Cell experiments confirmed that lactoalbumin-linoleic acid complex had a good anti-tumor effect. This study provides a basis for the development of a novel anti-tumor lactoalbumin-linoleic acid complex in terms of its structure and function.
【作者單位】: 北京工商大學食品學院;國家糧食局科學研究院;中國農(nóng)業(yè)大學教育部-北京市共建功能乳品重點實驗室;
【基金】:國家科技支撐計劃項目(2012BAD12B08) 國家國際科技合作專項(2011DFA32550)資助
【分類號】:R914;R96
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