Abstract
This chapter describes the cloning, expression, and purification of human Hsp10, also known as chaperonin-10, cpn10. The function of Hsp10 is to bind Hsp60 (also known as chaperonin-60) cpn60, in the presence of ATP, thereby promoting a conformational change in Hsp60 and enclosing the protein substrate within the complex (1). ATP hydrolysis by Hsp60 destabilizes the Hsp60/ Hsp10 complex, allowing it to dissociate and release the substrate protein. Crystal structures of the Escherichia coli (2), Mycobacterium leprae (3), and human Hsp10s (Hunt, et al., unpublished results) have been determined, and the architecture of the E. coli Hsp60/Hsp10 complex has been described by cryo-electron microscopy (4) and X-ray crystallography (5). Subunits of both Hsp10 and Hsp60 are arranged in sevenfold symmetric rings. Hsp10 is composed of 7 10-kDa subunits, and Hsp60 is composed of fourteen 60 kDa subunits.
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© 2000 Humana Press Inc.
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Kalaya Steede, N., Guidry, J.J., Landry, S.J. (2000). Preparation of Recombinant Human Hsp10. In: Schneider, C. (eds) Chaperonin Protocols. Methods in Molecular Biology, vol 140. Humana, Totowa, NJ. https://doi.org/10.1385/1-59259-061-6:145
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DOI: https://doi.org/10.1385/1-59259-061-6:145
Publisher Name: Humana, Totowa, NJ
Print ISBN: 978-0-89603-739-7
Online ISBN: 978-1-59259-061-2
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