利用尺寸排阻色谱法研究蛋白质的变性

通过比较蛋白质变性时的色谱行为和生物物理特性，提出利用尺寸排阻色谱法研究蛋白质变性时的构象变化，根据色谱参数中保留时间，比较蛋白质变性时体积的相对变化，利用色谱峰数，确定形成变体的数目，根据峰形和峰数的变化，描述蛋白质的伸展程度，利用不同波长下峰高的变化，推断蛋白质变性芳香族基酸残基的暴露情况，利用建立的尺寸排阻色谱观察了液体和固液α－淀粉酶在低温下放置时的变性情况，讨论了变性时间和变性温度对蛋白

Studies on Protein Denaturation by Sizc-exclusion Chromatography

A size-excltlsion chromatography able to study conformational change during protein denaturation was proposed by comparing the correlations between protein biophysicalproperties and chromatographic behaviors. The relatiove volume of denatured protein may becompared by e-exclusion chromatography in term o fchanges in retention time. The numberof forming denatured species can be determined by chromatographic peak number. The expansion extent of protein may be described using changes in peak shape and peak number.The exposure of aromatic amino acid residues in denatured proteins can also be deduced from peak height under different wave lengths. We utilized the extablished size-exclusion chromatography to examine the denatured aspects for liquid and solid a-amylase upon long term storage under lower temperature, to discuss the influence of denaturation time and denaturation temperature on unfolding behavior of proteins