Triton™ X-100在UV-Vis光谱中的行为类似于含酪氨酸和不含色氨酸的蛋白质

IF 1.4 Q3 CHEMISTRY, MULTIDISCIPLINARY
H. Nedaei, A. Saboury
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引用次数: 0

摘要

作为一种已知的非离子变性洗涤剂和乳化剂,Triton™ X-100经常用于各种生物化学研究,包括用于溶解膜蛋白的膜蛋白复合物的分离,作为裂解缓冲液成分的周质蛋白提取过程,作为渗透试剂的间接免疫荧光染色和流式细胞术等™ X-100的最佳pH范围为6.0-8.0,对紫外光有显著的吸收。在本项目中,我们展示了海卫一的吸收光谱™ X-100,当溶解在1X磷酸盐缓冲盐水中时,与α-突触核蛋白类似,因为这些蛋白质缺乏色氨酸,但含有酪氨酸作为其主要紫外线吸收剂。这些结果表明,无论何时使用Triton™ X-100用于提取膜和周质蛋白是不可避免的,在通过UV-Vis光谱对蛋白质进行表征之前清除它,特别是使用比尔-朗伯定律测定其浓度是必要的。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Triton™ X-100 Behaves Similarly to Tyrosine-Containing and Tryptophan-Free Proteins in UV-Vis Spectroscopy
As a known non-ionic, -denaturing detergent and emulsifier, Triton™ X-100 is often used in various biochemical studies including in the isolation of membrane-protein complexes for solubilizing membrane proteins, in the process of periplasmic protein extraction as a component of the lysis buffer, in both indirect immunofluorescence staining and flow cytometry as a permeabilization reagent, etc. It has been shown that the diluted solution of Triton™ X-100 with the optimal pH range of 6.0-8.0 has a significant absorption of UV light. In the present project, we show that the absorption spectrum of Triton™ X-100, when dissolved in 1X phosphate-buffered saline, is similar to that of α-synuclein, as a representative of those proteins lack tryptophan but contain tyrosine as their main UV absorber. These results show that whenever the use of Triton™ X-100 for extracting membrane and periplasmic proteins is inevitable, scavenging it before the characterization of the proteins by UV-Vis spectroscopy, especially the determination of their concentration using Beer-Lambert Law, would be necessary.
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来源期刊
Physical Chemistry Research
Physical Chemistry Research CHEMISTRY, MULTIDISCIPLINARY-
CiteScore
2.70
自引率
8.30%
发文量
18
期刊介绍: The motivation for this new journal is the tremendous increasing of useful articles in the field of Physical Chemistry and the related subjects in recent years, and the need of communication between Physical Chemists, Physicists and Biophysicists. We attempt to establish this fruitful communication and quick publication. High quality original papers in English dealing with experimental, theoretical and applied research related to physics and chemistry are welcomed. This journal accepts your report for publication as a regular article, review, and Letter. Review articles discussing specific areas of physical chemistry of current chemical or physical importance are also published. Subjects of Interest: Thermodynamics, Statistical Mechanics, Statistical Thermodynamics, Molecular Spectroscopy, Quantum Chemistry, Computational Chemistry, Physical Chemistry of Life Sciences, Surface Chemistry, Catalysis, Physical Chemistry of Electrochemistry, Kinetics, Nanochemistry and Nanophysics, Liquid Crystals, Ionic Liquid, Photochemistry, Experimental article of Physical chemistry. Mathematical Chemistry.
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