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Protein Science (2004), 13:134-144. Published by Cold Spring Harbor Laboratory Press. Copyright © 2004 The Protein Society
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Expression and biochemical characterization of two small heat shock proteins from the thermoacidophilic crenarchaeon Sulfolobus tokodaii strain 7

Keisuke Usui1, Noriyuki Ishii2, Yutaka Kawarabayasi3 and Masafumi Yohda1

1 Department of Biotechnology and Life Sciences, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588, Japan
2 Biological Information Research Center and
3 Research Center for Glycoscience, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki 305-8566, Japan

Reprint requests to: Masafumi Yohda, Department of Biotechnology and Life Sciences, Tokyo University of Agriculture and Technology, 2-24-16, Naka-cho, Koganei, Tokyo 184-8588, Japan; e-mail: yohda{at}cc.tuat.ac.jp; fax: 81-42-388-7479.

We expressed and characterized two sHsps, StHsp19.7 and StHsp14.0, from a thermoacidophilic crenarchaeon, Sulfolobus tokodaii strain 7. StHsp19.7 forms a filamentous structure consisting of spherical particles and lacks molecular chaperone activity. Fractionation of Sulfolobus extracts by size exclusion chromatography with immunoblotting indicates that StHsp19.7 exists as a filamentous structure in vivo. On the other hand, StHsp14.0 exists as a spherical oligomer like other sHsps. It showed molecular chaperone activity to protect thermophilic 3-isopropylmalate dehydrogenase (IPMDH) from thermal aggregation at 87°C. StHsp14.0 formed variable-sized complexes with denatured IPMDH at 90°C. Using StHsp14.0 labeled with fluorescence or biotin probe and magnetic separation, subunit exchanges between complexes were demonstrated. This is the first report on the filament formation of sHsp and also the high molecular chaperone activity of thermophilic archaeal sHsps.

Keywords: small heat shock protein; archaea; thermoacidophile; chaperone activity; oligomerization

Abbreviations: sHsp, small heat shock protein • IPMDH, 3-isopropylmalate dehydrogenase from Thermus thermophilus HB8 • SDS-PAGE, sodium dodecyl sulfate–polyacrylamide gel electrophoresis • CS, citrate synthase from porcine heart


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[Abstract] [Full Text] [PDF]


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