Study on the Characteristics of α-Chymotrypsin Folding Intermediates by Hydrophobic Interaction Chromatography (HIC) | Chapter 2 | Current Advances in Chemistry and Biochemistry Vol. 7

The study of protein folding intermediates is crucial for understanding the folding mechanism of denatured proteins and enhancing protein folding efficiency. A new methodology to define the intermediate of urea-denatured -chymotrypsin (-Chy) was established in this study by using some of the linear parameters of the stoichiometric displacement theory of retention of solute (SDT-R) of hydrophobic interaction chromatography (HIC). As urea concentration (Curea) fluctuates, the contact surface region (Z, S), affinity (logI), and character of interaction force (j) of the -Chy to the stationary phase of HIC (STHIC) between the intermediate (M) and native (N) states were shown to be considerably different. With modifications in Curea, a linear relationship between logI and Z was discovered only for its N state, not for its M state, implying that the interaction force between -Chy in N state and the STHIC is non-selective in N state but selective in M state. In addition, the magnitude of both logI and Z measured in the M state is only a fifth of that in the N state. To discriminate between proteins in the N and M states, all three parameters were used. This finding could be used to distinguish any non-functional protein with a correct three- or four-dimensional molecular structure from their stable M state of any kind of protein, and/or other proteins, in proteome research, protein separation, and a thorough understanding of the intrinsic rule of protein folding in molecular biology.

Author (s) Details

Congyu Ke
College of Chemistry and Chemical Engineering, Xi’an Shiyou University, Xi,’an 710065, China.

Wei Tuo
Schol of Foreign Languages, Xi’an Shiyou University, Xi’an 710065, China.

Wujuan Sun
College of Chemistry and Chemical Engineering, Xi’an Shiyou University, Xi,’an 710065, China.

Jianjun Li
Institute of Modern Separation Science, Shaanxi Key Laboratory of Modern Separation Science, Key Laboratory of Synthetic and Natural Functional Molecule Chemistry of Ministry of Education, Northwest University, 710069 Xi’an, P.R. China.

Zhenling Liu
Xinxiang Medical College, Xinxiang, 453003, Henan Province, P.R. China.

Xindu Geng
Institute of Modern Separation Science, Shaanxi Key Laboratory of Modern Separation Science, Key Laboratory of Synthetic and Natural Functional Molecule Chemistry of Ministry of Education, Northwest University, 710069 Xi’an, P.R. China.

View Book :- https://stm.bookpi.org/CACB-V7/article/view/1188