Adsorption behavior of human serum albumin on ATR crystal studied by in situ ATR/FTIR spectroscopy and two-dimensional correlation analysis

被引:19
作者
Huang, He [1 ]
Xie, Jing [1 ,2 ]
Chen, Hong [1 ]
机构
[1] Soochow Univ, Coll Chem Chem Engn & Mat Sci, Suzhou 215123, Peoples R China
[2] Wuhan Univ Technol, Sch Mat Sci & Engn, Wuhan 430070, Peoples R China
基金
中国国家自然科学基金;
关键词
CORRELATION INFRARED-SPECTROSCOPY; PROTEIN SECONDARY STRUCTURE; STRUCTURAL-CHANGES; AQUEOUS-SOLUTIONS; SEQUENTIAL ORDER; POLYMER BLENDS; RESOLUTION; INTERFACE;
D O I
10.1039/c0an00890g
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
The time-dependent adsorption behavior of human serum albumin (HSA) onto an ATR (ZnSe) crystal was investigated by two-dimensional (2D) correlation analysis and in situ ATR-FTIR spectroscopy following the secondary structural changes in the amide I region. The two major advantages of the generalized 2D correlation spectroscopy were first tested. New extra bands have been resolved by 2D correlation analysis, but they are either artifacts or a result of uncertainty on band position in generalized 2D correlation spectroscopy. The sequence of the intensity variations of the three sub-bands under the amide I band profile deduced from the 'sequential order' rules is contradictory to the experimental observation, which supports our argument on the 'sequential order' rules in generalized 2D correlation spectroscopy (H. Huang, Anal. Chem., 2007, 79, 8281-8292). Subsequent detailed analysis on the in situ ATR-IR spectra shows that the adsorption process of HSA on the ATR (ZnSe) crystal in aqueous solutions can be divided into three stages: no obvious conformational transitions in the first 25 min of adsorption of HSA molecules; large structural rearrangement from a-helix to random coil and short extended chain structures in a fully cooperative way from 25 to 50 min of adsorption; and further slight conformational transformation of short extended chain and turn structures into random coil with no sequential order after 50 min of adsorption.
引用
收藏
页码:1747 / 1752
页数:6
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