NMR structures of two designed proteins with high sequence identity but different fold and function.

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TitleNMR structures of two designed proteins with high sequence identity but different fold and function.
Publication TypeJournal Article
Year of Publication2008
AuthorsHe, Y, Chen, Y, Alexander, P, Bryan, PN, Orban, J
JournalProc Natl Acad Sci U S A
Date Published2008 Sep 23
KeywordsAmino Acid Sequence, Bacterial Proteins, Humans, Models, Molecular, Molecular Sequence Data, Mutant Proteins, Nuclear Magnetic Resonance, Biomolecular, Protein Folding, Protein Structure, Secondary, Protein Structure, Tertiary, Sequence Homology, Amino Acid, Solvents, Streptococcus, Structure-Activity Relationship

How protein sequence codes for 3D structure remains a fundamental question in biology. One approach to understanding the folding code is to design a pair of proteins with maximal sequence identity but retaining different folds. Therefore, the nonidentities must be responsible for determining which fold topology prevails and constitute a fold-specific folding code. We recently designed two proteins, G(A)88 and G(B)88, with 88% sequence identity but different folds and functions [Alexander et al. (2007) Proc Natl Acad Sci USA 104:11963-11968]. Here, we describe the detailed 3D structures of these proteins determined in solution by NMR spectroscopy. Despite a large number of mutations taking the sequence identity level from 16 to 88%, G(A)88 and G(B)88 maintain their distinct wild-type 3-alpha and alpha/beta folds, respectively. To our knowledge, the 3D-structure determination of two monomeric proteins with such high sequence identity but different fold topology is unprecedented. The geometries of the seven nonidentical residues (of 56 total) provide insights into the structural basis for switching between 3-alpha and alpha/beta conformations. Further mutation of a subset of these nonidentities, guided by the G(A)88 and G(B)88 structures, leads to proteins with even higher levels of sequence identity (95%) and different folds. Thus, conformational switching to an alternative monomeric fold of comparable stability can be effected with just a handful of mutations in a small protein. This result has implications for understanding not only the folding code but also the evolution of new folds.

Alternate JournalProc. Natl. Acad. Sci. U.S.A.
PubMed ID18796611
PubMed Central IDPMC2567172
Grant ListGM62154 / GM / NIGMS NIH HHS / United States