Biochemical and functional characterization of charge-defined subfractions of high-density lipoprotein from normal adults.
Biochemical and functional characterization of charge-defined subfractions of high-density lipoprotein from normal adults.
复制标题
DOI：
10.1021/ac402516u
复制
发表时间：
2013-12-03
影响因子：
7.4
通讯作者：
Yang, Chao-Yuh
中科院分区：
文献类型：
--
作者：
Hsieh, Ju-Yi;Chang, Chiz-Tzung;Huang, Max T.;Chang, Chia-Ming;Chen, Chia-Ying;Shen, Ming-Yi;Liao, Hsin-Yi;Wang, Guei-Jane;Chen, Chu-Huang;Chen, Chao-Jung;Yang, Chao-Yuh
高密度脂蛋白（HDL）被认为具有动脉粥样硬化保护作用，因为它具有抗氧化和抗炎作用，并在逆向胆固醇运输中起重要作用。在本文中，我们概述了一种研究高密度脂蛋白异质性的新方法。采用阴离子交换色谱法，将6名健康人的HDL分离成5个带电荷的亚段（H1 ~ H5），并对各亚段的组成和生物活性进行了评价。十二烷基硫酸钠聚丙烯酰胺凝胶电泳分析显示，载脂蛋白（apo） AI和apoAII均存在于5个亚段中；apoi仅在H1中存在；apoCIII和apoE在H4和H5中含量最高。hdl相关的抗氧化酶如卵磷脂-胆固醇酰基转移酶、脂蛋白相关磷脂酶A2和对氧磷酶1在H4和H5中最为丰富。采用基质辅助激光解吸飞行时间质谱法分析各亚段的脂蛋白同工型。为了定量HDL亚组分中的其他蛋白质，我们使用等压标签进行相对和绝对定量方法，然后使用纳流液相色谱-串联质谱分析。检测到的抗氧化蛋白以H4和H5居多。各亚组分诱导巨噬细胞胆固醇外排的能力随着HDL电负性的增加而增强，但H5促进外排活性最小。总之，阴离子交换色谱法是一种很有吸引力的方法，可以将高密度脂蛋白分离成具有不同脂蛋白组成和生物活性的亚组分。通过比较这些亚组分的特性，有可能发现在疾病中起作用的hdl特异性蛋白质。
High-density lipoprotein (HDL) is regarded as atheroprotective because it provides antioxidant and anti-inflammatory benefits and plays an important role in reverse cholesterol transport. In this paper, we outline a novel methodology for studying the heterogeneity of HDL. Using anion-exchange chromatography, we separated HDL from 6 healthy individuals into 5 subfractions (H1 through H5) with increasing charge and evaluated the composition and biologic activities of each subfraction. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis showed that apolipoprotein (apo) AI and apoAII were present in all 5 subfractions; apoCI was present only in H1; and apoCIII and apoE were most abundantly present in H4 and H5. HDL-associated antioxidant enzymes such as lecithin-cholesterol acyltransferase, lipoprotein-associated phospholipase A2, and paraoxonase 1 were most abundant in H4 and H5. Lipoprotein isoforms were analyzed in each subfraction by using matrix-assisted laser desorption–time of flight mass spectrometry. To quantify other proteins in the HDL subfractions, we used the isobaric tags for relative and absolute quantitation approach followed by nanoflow liquid chromatography–tandem mass spectrometry analysis. Most antioxidant proteins detected were found in H4 and H5. The ability of each subfraction to induce cholesterol efflux from macrophages increased with increasing HDL electronegativity, with the exception of H5, which promoted the least efflux activity. In conclusion, anion-exchange chromatography is an attractive method for separating HDL into subfractions with distinct lipoprotein compositions and biologic activities. By comparing the properties of these subfractions, it may be possible to uncover HDL-specific proteins that play a role in disease.
登录
查看更多内容
影响因子：
6.2
作者：
Chen, Chao-Jung;Lai, Chien-Chen;Tsai, Fuu-Jen
通讯作者：
Tsai, Fuu-Jen
影响因子：
5
作者：
Chang, Chiz-Tzung;Liao, Hsin-Yi;Chen, Chao-Jung
通讯作者：
Chen, Chao-Jung
DOI：
10.1016/j.bbrc.2005.04.129
发表时间：
2005-07-01
影响因子：
3.1
作者：
Farwig, ZN;McNeal, CJ;Macfarlane, RD
通讯作者：
Macfarlane, RD
影响因子：
120.7
作者：
CASTELLI, WP;GARRISON, RJ;KANNEL, WB
通讯作者：
KANNEL, WB
影响因子：
5
作者：
FOURNIER, N;MYARA, I;MOATTI, N
通讯作者：
MOATTI, N