Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
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Nucleotide Triphosphates Inhibit the Degradation of Unfolded Proteins by HslV Peptidase

  • Lee, Jung Wook (School of Biological Sciences, Seoul National University) ;
  • Park, Eunyong (School of Biological Sciences, Seoul National University) ;
  • Bang, Oksun (School of Biological Sciences, Seoul National University) ;
  • Eom, Soo-Hyun (Department of Biological Sciences, Gwangju Institute of Science and Technology) ;
  • Cheong, Gang-Won (Division of Applied Life Sciences and Environmental Biotechnology National Core Research Center, Gyeongsang National University) ;
  • Chung, Chin Ha (School of Biological Sciences, Seoul National University) ;
  • Seol, Jae Hong (School of Biological Sciences, Seoul National University)
  • Received : 2007.01.23
  • Accepted : 2007.01.29
  • Published : 2007.04.30
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Abstract

Escherichia coli HslVU is an ATP-dependent protease consisting of two heat shock proteins, the HslU ATPase and HslV peptidase. In the reconstituted enzyme, HslU stimulates the proteolytic activity of HslV by one to two orders of magnitude, while HslV increases the rate of ATP hydrolysis by HslU several-fold. Here we show that HslV alone can efficiently degrade certain unfolded proteins, such as unfolded lactalbumin and lysozyme prepared by complete reduction of disulfide bonds, but not their native forms. Furthermore, HslV alone cleaved a lactalbumin fragment sandwiched by two thioredoxin molecules, indicating that it can hydrolyze the internal peptide bonds of lactalbumin. Surprisingly, ATP inhibited the degradation of unfolded proteins by HslV. This inhibitory effect of ATP was markedly diminished by substitution of the Arg86 residue located in the apical pore of HslV with Gly, suggesting that interaction of ATP with the Arg residue blocks access of unfolded proteins to the proteolytic chamber of HslV. These results suggest that uncomplexed HslV is inactive under normal conditions, but may can degrade unfolded proteins when the ATP level is low, as it is during carbon starvation.

Keywords

Acknowledgement

Supported by : Korea Science and Engineering Foundation

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