Int. J. Mol. Sci. 2009, 10(2), 616-628; doi:10.3390/ijms10020616
Article

A New Approach for Characterizing the Intermediate Feature of α-Chymotrypsin Refolding by Hydrophobic Interaction Chromatography

1 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 2 Xinxiang Medical College, Xinxiang, 453003, Henan Province, P.R. China
* Author to whom correspondence should be addressed.
Received: 31 October 2008; in revised form: 15 February 2009 / Accepted: 17 February 2009 / Published: 18 February 2009
(This article belongs to the Special Issue Protein Folding)
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Abstract: A new approach for characterizing the intermediate of urea-denatured α-chymotrypsin (α-Chy) by hydrophobic interaction chromatography (HIC) is presented. The contact surface region (Z, S), affinity (logI), and the character of interaction force (j) of the α-Chy to the stationary phase of HIC (STHIC) between the intermediate (M) and native (N) states were found to be quite different as urea concentration (Curea) changes. With the changes in Curea, a linear relationship between logI and Z was found to exist only for its N state, not for M state, indicating the interaction force between α-Chy in N state to the STHIC to be non-selective, but selective one for its M state. Also, the measured magnitude of both logI and Z in M state is only a fifth of that in N state. All three parameters were employed to distinguish protein in the N state from that in the M state. It would be expected that this result could be employed to distinguish any kind of non-functional protein having correct three-, or four-dimensional molecular structure from their stable M state of any kinds of proteins, and/or other proteins in proteome investigation, separation process of protein, and intensively understanding the intrinsic rule of protein folding in molecular biology.
Keywords: Protein folding; protein drugs; misfolding; intermediates; protein folding liquid chromatography; characterization; hydrophobic interaction chromatography; α-Chymotrypsin; stoichiometric displacement theory

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MDPI and ACS Style

Ke, C.; Li, J.; Liu, Z.; Geng, X. A New Approach for Characterizing the Intermediate Feature of α-Chymotrypsin Refolding by Hydrophobic Interaction Chromatography. Int. J. Mol. Sci. 2009, 10, 616-628.

AMA Style

Ke C, Li J, Liu Z, Geng X. A New Approach for Characterizing the Intermediate Feature of α-Chymotrypsin Refolding by Hydrophobic Interaction Chromatography. International Journal of Molecular Sciences. 2009; 10(2):616-628.

Chicago/Turabian Style

Ke, Congyu; Li, Jianjun; Liu, Zhenling; Geng, Xindu. 2009. "A New Approach for Characterizing the Intermediate Feature of α-Chymotrypsin Refolding by Hydrophobic Interaction Chromatography." Int. J. Mol. Sci. 10, no. 2: 616-628.

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