Similarities between the spectrin SH3 domain denatured state and its folding transition state.
We have expanded our description of the energy landscape for folding of the SH3 domain of chicken alpha-spectrin by a detailed structural characterization of its denatured state ensemble (DSE). This DSE is significantly populated under mildly acidic conditions in equilibrium with the folded state. Evidence from heteronuclear nuclear magnetic resonance (NMR) experiments on (2)H, (15)N-labeled protein suggests the presence of conformers whose residual structure bears some resemblence to the structure of the folding transition state of this protein. NMR analysis in a mutant with an engineered, non-native alpha-helical tendency shows a significant amount of local non-native structure in the mutant, while the overall characteristics of the DSE are unchanged. Comparison with recent theoretical predictions of SH3 domain folding reactions reveals an interesting correlation with the predicted early events. Based on these results and recent data from other systems, we propose that the DSE of a protein will resemble the intermediate or transition state of its nearest rate-limiting step, as a consequence of simple energetic and kinetic principles.
Kortemme
T
T
European Molecular Biology Laboratory (EMBL), Meyerhofstrasse 1, Heidelberg, D-6917, Germany. kortemme@u.washington.edu
Kelly
M J
MJ
Kay
L E
LE
Forman-Kay
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Serrano
L
L
D000143:Acids / Q000494:pharmacology; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D002645:Chickens; D003903:Deuterium / Q000378:metabolism; D029721:Drosophila Proteins; D006863:Hydrogen-Ion Concentration; D019476:Insect Proteins / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D019906:Nuclear Magnetic Resonance, Biomolecular; D011489:Protein Denaturation / Q000187:drug effects; D017510:Protein Folding; D020673:Protein Renaturation; D017433:Protein Structure, Secondary / Q000187:drug effects; D011522:Protons; D013049:Spectrin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D013816:Thermodynamics; D018909:src Homology Domains / Q000187:drug effects / Q000235:genetics
D000143:Acids; D029721:Drosophila Proteins; D019476:Insect Proteins; D011522:Protons; C080530:drk protein, Drosophila; D013049:Spectrin; D003903:Deuterium