Complex folding kinetics of a multidomain protein

Complex folding kinetics of a multidomain protein
Engineering Village 2
? 2006 Elsevier Inc
Accession number: 8915524

Title: Complex folding kinetics of a multidomain protein

Authors: Batey, S.1 ; Scott, K.A.1 ; Clarke, J.1

Author affiliation: 1 Dept. of Chem., Cambridge Univ., UK

Serial title: Biophysical Journal

Abbreviated serial title: Biophys. J. (USA)

Volume: 90

Issue: 6

Publication date: 15 March 2006

Pages: 2120-30

Language: English

ISSN: 0006-3495


Document type: Journal article (JA)

Publisher: Biophys. Soc

Country of publication: USA

Material Identity Number: B154-2006-006

Abstract: Spectrin domains are three-helix bundles, commonly found in large tandem arrays. Equilibrium studies have shown that spectrin domains are significantly stabilized by their neighbors. In this work we show that domain:domain interactions can also have profound effects on their kinetic behavior. We have studied the folding of a tandem pair of spectrin domains (R1617) using a combination of single- and double-jump stopped flow experiments (monitoring folding by both circular dichroism and fluorescence). Mutant proteins were also used to investigate the complex folding kinetics. We find that, although the domains fold and unfold individually, there is a single rate-determining step for both folding and unfolding of the protein. This is consistent with the equilibrium observation of cooperative folding of the entire two-domain protein. The results may have important biological implications. Not only will the protein fold more efficiently during cotranslational folding, but the ability of the multidomain protein to withstand thermal unfolding in the cell will be dramatically increased. This study suggests that caution has to be exercised when extrapolating from single domains to larger proteins with a number of independently folding modules arranged in tandem. The multidomain protein spectrin is certainly more than "the sum of its parts"

Number of references: 33

Inspec controlled terms: biochemistry - cellular biophysics - circular dichroism - fluorescence - molecular biophysics - proteins

Uncontrolled terms: complex folding kinetics - multidomain protein - spectrin domains - domain-domain interactions - kinetic behavior - single-jump stopped flow experiment - double-jump stopped flow experiment - circular dichroism - fluorescence - mutant proteins - cotranslational folding - thermal unfolding - cell

Inspec classification codes: A8715D Physical chemistry of biomolecular solutions; condensed states - A8715B Biomolecular structure, configuration, conformation, and active sites - A8715M Interactions with radiations at the biomolecular level - A8725F Physics of subcellular structures - A3620C Macromolecular conformation (statistics and dynamics) - A3345D Optical activity, optical rotation, circular dichroism in molecules - A3350D Molecular fluorescence and phosphorescence spectra

Treatment: Experimental (EXP)

Discipline: Physics (A)

DOI: 10.1529/biophysj.105.072710

Database: Inspec

Copyright 2006, The Institution of Engineering and Technology
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