识别老年人类镜片中紫外线滤光片与蛋白质的附着位点

J.A Aquilina, R.J.W Truscott
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引用次数: 35

摘要

最近的研究结果表明,色氨酸衍生的紫外线滤光剂对蛋白质的共价修饰可能解释了人类晶状体的年龄依赖性颜色,并在年龄相关性白内障中发挥作用。然而,紫外滤光片附着在晶状体结晶蛋白上的位置尚未确定。本研究利用一个预测紫外过滤器修饰的色氨酸肽质量的数据库来定位紫外过滤器附着的位点。从旧的正常晶状体中分离蛋白质,并在pH 6下用胰蛋白酶消化,以保持修饰位点的完整性。肽段采用高效液相色谱法分离,质谱法表征。消化的主要彩色和荧光峰被发现与含半胱氨酸的肽相对应,其中侧链的硫原子与主要的紫外线过滤器化合物3-羟基犬尿氨酸葡萄糖苷相连。其中3个肽来自γ - s -晶体蛋白,1个来自β b1 -晶体蛋白。这些结果表明,预测的质量数据库可用于人类晶状体结晶蛋白紫外滤光器修饰位点的识别。此外,这项工作首次证明了紫外线滤光片在体内与晶状体蛋白上的特定残基结合,并表明巯基可能是紫外线滤光片附着的重要位点。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Identifying sites of attachment of UV filters to proteins in older human lenses

Recent results indicate that covalent modification of proteins by tryptophan-derived UV filters may explain the age-dependent coloration of human lenses, and play a role in age-related cataract. The sites of attachment of the UV filters to the lens crystallins, however, have not been determined. This study utilized a database of predicted masses of UV filter-modified tryptic peptides to target sites of UV filter attachment. Proteins were isolated from old normal lenses and digested with trypsin at pH 6, in order to preserve the integrity of the sites of modification. Peptides were separated by high-performance liquid chromatography and characterized by mass spectrometry. Major colored and fluorescent peaks in the digest were found to correspond to cysteine-containing peptides in which the sulfur atom of the sidechain was linked to the major UV filter compound, 3-hydroxykynurenine glucoside. Three of the peptides originated from γS-crystallin and one from βB1-crystallin. These results show that a predicted mass database can be used to facilitate the identification of sites of UV filter modification in human lens crystallins. Furthermore, this work represents the first evidence that UV filters bind to specific residues on lens proteins in vivo, and suggests that sulfhydryl groups may be important sites for the attachment of UV filters.

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