Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
권호 표지

Biochemical Analysis of a Cytosolic Small Heat Shock Protein, NtHSP18.3, from Nicotiana tabacum

  • Yu, Ji Hee (Institute of Molecular Biology and Genetics, Seoul National University) ;
  • Kim, Keun Pill (Institute of Molecular Biology and Genetics, Seoul National University) ;
  • Park, Soo Min (Division of Biological Sciences, University of California at Davis) ;
  • Hong, Choo Bong (Institute of Molecular Biology and Genetics, Seoul National University)
  • Received : 2004.10.18
  • Accepted : 2005.01.31
  • Published : 2005.06.30
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Abstract

Small heat shock proteins (sHSPs) are widely distributed, and their function and diversity of structure have been much studied in the field of molecular chaperones. In plants, which frequently have to cope with hostile environments, sHSPs are much more abundant and diverse than in other forms of life. In response to high temperature stress, sHSPs of more than twenty kinds can make up more than 1% of soluble plant proteins. We isolated a genomic clone, NtHSP18.3, from Nicotiana tabacum that encodes the complete open reading frame of a cytosolic class I small heat shock protein. To investigate the function of NtHSP18.3 in vitro, it was overproduced in Escherichia coli and purified. The purified NtHSP18.3 had typical molecular chaperone activity as it protected citrate synthase and luciferase from high temperature-induced aggregation. When E. coli celluar proteins were incubated with NtHSP18.3, a large proportion of the proteins remained soluble at temperatures as high as $70^{\circ}C$. Native gel analysis suggested that NtHSP18.3 is a dodecameric oligomer as the form present and showing molecular chaperone activity at the condition tested. Binding of bis-ANS to the oligomers of NtHSP18.3 indicated that exposure of their hydrophobic surfaces increased as the temperature was raised. Taken together, our data suggested that NtHSP18.3 is a molecular chaperone that functions as a dodecameric complex and possibly in a temperature-induced manner.

Keywords

Acknowledgement

Supported by : Crop Functional Genomics Center, Ministry of Science and Technology of Korea

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