The sequences of small proteins are not extensively optimized for rapid folding by natural selection.
The thermodynamic stabilities of small protein domains are clearly subject to natural selection, but it is less clear whether the rapid folding rates typically observed for such proteins are consequences of direct evolutionary optimization or reflect intrinsic physical properties of the polypeptide chain. This issue can be investigated by comparing the folding rates of laboratory-generated protein sequences to those of naturally occurring sequences provided that the method by which the sequences are generated has no kinetic bias. Herein we report the folding thermodynamics and kinetics of 12 heavily mutated variants of the small IgG binding domain of protein L retrieved from high-complexity combinatorial libraries by using a phage-display selection for proper folding that does not discriminate between rapidly and slowly folding proteins. Although the stabilities of all variants were decreased, many of the variants fold faster than wild type. Taken together with similar results for the src homology 3 domain, this observation suggests that the sequences of small proteins have not been extensively optimized for rapid folding; instead, rapid folding appears to be a consequence of selection for stability.
Kim
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DE
Department of Biochemistry, University of Washington, Seattle, WA 98195, USA.
Gu
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Baker
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D001426:Bacterial Proteins / Q000737:chemistry*; D007700:Kinetics; D008970:Molecular Weight; D010462:Peptostreptococcus; D017510:Protein Folding; D011994:Recombinant Proteins; D013329:Structure-Activity Relationship; D013816:Thermodynamics
D001426:Bacterial Proteins; C055047:Ig L-binding protein, Peptostreptococcus; D011994:Recombinant Proteins