Dehydroalanine crosslinks in human lens
Dehydroalanine crosslinks in human lens
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DOI：
10.1016/j.exer.2004.06.026
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发表时间：
2004-10-01
影响因子：
3.4
通讯作者：
Hu, F
中科院分区：
文献类型：
--
作者：
Linetsky, M;Hill, JMW;Hu, F
本研究旨在开发一种纯化和检测晶状体组织中组氨酸丙氨酸、羊毛硫氨酸和赖氨酸丙氨酸的方法。使用阴离子交换色谱法对白内障和老化的人晶状体蛋白进行水解和分级。含有大量脱氢丙氨酸交联的级分用丹磺酰氯衍生化，然后通过RP-HPLC进行分离和定量。本研究中纯化和定量的所有三种物质的光谱和色谱特性与其合成对应物相同。组氨酸丙氨酸和羊毛硫氨酸是水溶性和水不溶性晶状体蛋白中最丰富的脱氢丙氨酸交联。印度白内障晶状体水溶性蛋白中的组氨酸丙氨酸水平比正常晶状体水溶性蛋白高 6.2 倍（1(.)68 +/- 0(.)75 vs 0(.)26 +/- 0(.)06 nmol/mg 蛋白；p < 0(.)001）。在水不溶性蛋白质中，白内障晶状体中的蛋白质含量比正常晶状体高 2 (.)2 倍（1(.)59 +/- 0(.)76 vs 0(.)73 +/- 0(.)17 nmol/mg 蛋白质；p < 0.01）。与非白内障晶状体相比，白内障晶状体的水不溶性蛋白质中的羊毛硫氨酸水平显着较高（2(.)5 +/- 1(.)68 vs 0(.)95 +/- 0(.)08 nmol/mg 蛋白质；p < 0(.)03）。与组氨酸丙氨酸不同，这种交联似乎在水溶性晶状体蛋白中以相对较高的浓度积累。其浓度比相同蛋白质的组氨酸丙氨酸高 9 倍（0(.)26 +/- 0(.)06 HAL vs 2(.)34 +/- 0(.)76 LAN nmol/mg 蛋白质；p < 0(.)0004）。赖氨酸丙氨酸的浓度在皮摩尔范围内并且仅存在于白内障晶状体蛋白中。 (C) 2004 Elsevier Ltd. 保留所有权利。
This study was conducted to develop a methodology for the purification and detection of histidinoalanine, lanthionine and lysinoalanine in the lens tissue. Cataractous and aged human lens proteins were hydrolysed and fractionated by using anion-exchange chromatography. The fraction containing the bulk of dehydroalanine crosslinks was derivatized with dansyl chloride and then separated and quantified by means of RP-HPLC. The spectral and chromatographic properties of all three substances purified and quantified in this study were identical to those of their synthesized counterparts. Histidinoalanine and lanthionine were the most abundant dehydroalanine crosslinks in both water-soluble and water-insoluble lens proteins. Histidinoalanine levels in water-soluble proteins from the cataractous lenses of Indian origin were 6.2-fold higher than those in water-soluble proteins from normal lenses (1(.)68 +/- 0(.)75 vs 0(.)26 +/- 0(.)06 nmol/mg protein; p < 0(.)001). In water-insoluble proteins, they were 2 (.) 2-fold higher in cataractous lenses compared with normal lenses (1(.)59 +/- 0(.)76 vs 0(.)73 +/- 0(.)17 nmol/mg protein; p < 0.01). Lanthionine levels were significantly higher in water-insoluble proteins of cataractous lenses when compared to non-cataractous lenses (2(.)5 +/- 1(.)68 vs 0(.)95 +/- 0(.)08 nmol/mg protein; p < 0(.)03). Unlike histidinoalanine, this crosslink appears to accumulate in relatively high concentrations in water-soluble lens proteins; its concentration was 9-fold higher than histidinoalanine from the same proteins (0(.)26 +/- 0(.)06 HAL vs 2(.)34 +/- 0(.)76 LAN nmol/mg protein; p < 0(.)0004). The concentration of lysinoalanine was in the picomolar range and in cataractous lens proteins only. (C) 2004 Elsevier Ltd. All rights reserved.