REVERSED-PHASE LIQUID-CHROMATOGRAPHY AS A USEFUL PROBE OF HYDROPHOBIC INTERACTIONS INVOLVED IN PROTEIN-FOLDING AND PROTEIN STABILITY

被引:51
作者
HODGES, RS
ZHU, BY
ZHOU, NE
MANT, CT
机构
[1] UNIV ALBERTA,PROT ENGN NETWORK CTR EXCELLENCE,EDMONTON T6G 2H7,AB,CANADA
[2] UNIV ALBERTA,MRC,PROT STRUCT & FUNCT GRP,EDMONTON T6G 2H7,AB,CANADA
基金
英国医学研究理事会;
关键词
D O I
10.1016/0021-9673(94)80452-4
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
We have evaluated the potential of reversed-phase liquid chromatography (RPLC) as a probe of hydrophobic interactions involved in protein folding and stability. Our approach was to apply RPLC to a de novo designed model protein system, namely a two-stranded alpha-helical coiled coil. It was shown that the reversed-phase retention behaviour of various synthetic analogues of monomeric alpha-helices and dimeric coiled-coil structures correlated well with their stability in solution, as monitored by circular dichroism during guanidine hydrochloride and temperature denaturation studies. In addition, an explanation is offered as to why amphipathic coiled coils, an important structural motif in many biological systems, are more stable at low pH compared to physiological pH values. The results;of this study suggest that not only may RPLC prove to be a useful and rapid complementary technique for understanding protein interactions, but also the de novo designed coiled-coil model described here is an excellent model system for such studies.
引用
收藏
页码:3 / 15
页数:13
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