title: Complex folding kinetics of a multidomain protein.
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".
authors.affiliation: Department of Chemistry, MRC Centre for Protein Engineering, University of Cambridge, Cambridge CB2 1EW, United Kingdom.
authors.forename: Sarah
authors.initials: S
authors.lastname: Batey
authors.forename: Kathryn A
authors.initials: KA
authors.lastname: Scott
authors.forename: Jane
authors.initials: J
authors.lastname: Clarke
mesh_terms: D001665:Binding Sites; D003198:Computer Simulation; D007700:Kinetics; D008956:Models, Chemical; D008958:Models, Molecular; D011485:Protein Binding; D011487:Protein Conformation; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D013049:Spectrin / Q000737:chemistry* / Q000648:ultrastructure
chemical_list: D013049:Spectrin