Pressure equilibrium and jump study on unfolding of 23-kDa protein from spinach photosystem II

Cui Yan Tan, Chun He Xu, Jun Wong, Jian Ren Shen, Shinsuke Sakuma, Yasusi Yamamoto, Reinhard Lange, Claude Balny, Kang Cheng Ruan

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27 Citations (Scopus)

Abstract

Pressure-induced unfolding of 23-kDa protein from spinach photosystem Il has been systematically investigated at various experimental conditions. Thermodynamic equilibrium studies indicate that the protein is very sensitive to pressure. At 20°C and pH 5.5, 23-kDa protein shows a reversible two-state unfolding transition under pressure with a midpoint near 160 MPa, which is much lower than most natural proteins studied to date. The free energy (ΔG u) and volume change (ΔVu) for the unfolding are 5.9 kcal/mol and -160 ml/mol, respectively. It was found that NaCl and sucrose significantly stabilize the protein from unfolding and the stabilization is associated not only with an increase in ΔGu but also with a decrease in ΔVu. The pressure-jump studies of 23-kDa protein reveal a negative activation volume for unfolding (-66.2 ml/mol) and a positive activation volume for refolding (84.1 ml/mol), indicating that, in terms of system volume, the protein transition state lies between the folded and unfolded states. Examination of the temperature effect on the unfolding kinetics indicates that the thermal expansibility of the transition state and the unfolded state of 23-kDa protein are closer to each other and they are larger than that of the native state. The diverse pressure-refolding pathways of 23-kDa protein in some conditions were revealed in pressure-jump kinetics.

Original languageEnglish
Pages (from-to)1264-1275
Number of pages12
JournalBiophysical Journal
Volume88
Issue number2
DOIs
Publication statusPublished - Feb 2005

ASJC Scopus subject areas

  • Biophysics

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    Tan, C. Y., Xu, C. H., Wong, J., Shen, J. R., Sakuma, S., Yamamoto, Y., Lange, R., Balny, C., & Ruan, K. C. (2005). Pressure equilibrium and jump study on unfolding of 23-kDa protein from spinach photosystem II. Biophysical Journal, 88(2), 1264-1275. https://doi.org/10.1529/biophysj.104.050435