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The extremely slow-exchanging core and acid-denatured state of green fluorescent protein

Huang, J.-R. and Hsu, S. -T. D. and Christodoulou, J. and Jackson, S. E.. (2008) The extremely slow-exchanging core and acid-denatured state of green fluorescent protein. HFSP Journal, Vol. 2, H. 6. pp. 378-387.

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Official URL: http://edoc.unibas.ch/dok/A5249271

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Abstract

Green fluorescent protein (GFP) is a large protein with a complex eleven-stranded -barrel structure. Previous studies have shown that it has a complex energy landscape for folding on which there are several intermediate states and a denatured state with significant residual structure. Here, we use two different types of H/D exchange measurement and nuclear magnetic resonance (NMR) techniques to probe the energy landscape for folding of GFP in further detail. H/D exchange experiments were performed over a wide range of conditions including different concentrations of denaturant. Results show that the penetration model dominates the exchange mechanism, consistent with the known stability and slow unfolding kinetics of GFP. H/D exchange experiments at high pH establish that there is an extremely slow-exchanging superstable core of amide protons in GFP that are clustered and located in -strands 1, 2, 4, 5, and 6. These residues form part of a mini- -sheet which we propose constitutes a folding nucleus. Using a pulsed-labeling strategy, the acid-denatured state has been investigated and the residual structure observed in earlier studies shown to locate to -strands 1 and 3. There is some evidence that this residual structure is stabilized by a localized hydrophobic collapse of the polypeptide chain.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Structural Biology & Biophysics > Structural Biology (Grzesiek)
UniBasel Contributors:Huang, Jie-rong
Item Type:Article, refereed
Article Subtype:Research Article
Bibsysno:Link to catalogue
Publisher:HFSP Publishing
ISSN:1955-2068
Note:Publication type according to Uni Basel Research Database: Journal article
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Last Modified:22 Mar 2012 14:19
Deposited On:22 Mar 2012 13:16

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