An archaeal chaperonin-based reactor for renaturation of denatured proteins |
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Authors: | L Cerchia M Rossi A Guagliardi |
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Institution: | (1) Dipartimento di Chimica Organica e Biologica, Università degli Studi di Napoli, Via Mezzocannone 16, 80134, Napoli, Italy Tel. +39-81-7041276; Fax +39-81-5521217 e-mail: guaglia@unina.it, IT;(2) Istituto di Biochimica delle Proteine ed Enzimologia, C.N.R., Napoli, Italy, IT |
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Abstract: | We describe an original chaperonin-based reactor that yields folded and active proteins from denatured materials. We used
the 920-kDa chaperonin of the archaeon Sulfolobus solfataricus, which does not require any protein partner for its full activity and assists in vitro folding with low substrate specificity.
The reactor consists of an ultrafiltration cell equipped with a membrane that retains the chaperonin in a functional state
for folding in solution and permits the flowthrough of the folded substrates. By studying the ATP-dependent functional cycle
of the chaperonin, we were able to use the reactor for repeated refolding processes. The scale-up of the reactor is made possible
by the overproduction of chaperonin in Sulfolobus solfataricus cells that acquired thermotolerance upon appropriate heat shock.
Received: January 24, 1999 / Accepted: August 7, 1999 |
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Keywords: | Archaea Chaperonin Protein folding Heat shock Thermotolerance |
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