Mechanism and energy landscape of domain swapping in the B1 domain of protein G.

TitleMechanism and energy landscape of domain swapping in the B1 domain of protein G.
Publication TypeJournal Article
Year of Publication2008
AuthorsMalevanets, A., Sirota F. L., and Wodak S. J.
JournalJ Mol Biol
Volume382
Issue1
Pagination223-35
Date Published2008 Sep 26
ISSN1089-8638
KeywordsAmino Acid Sequence, Bacterial Proteins, Computer Simulation, Dimerization, Molecular Sequence Data, Motion, Mutant Proteins, Protein Binding, Protein Structure, Secondary, Protein Structure, Tertiary, Thermodynamics
Abstract

Three-dimensional domain swapping has emerged as a ubiquitous process for homo-oligomer formation in many unrelated proteins, but the molecular mechanism of this process is still poorly understood. Here we present a mechanism for the swapping reaction in the B1 domain of the immunoglobulin G binding protein from group G of Streptococcus (GB1). This is a particularly attractive system for investigating the swapping process, as the swapped dimer formed by the quadruple mutant (L5V/F30V/Y33F/A34F) of GB1 was recently shown to exist in equilibrium with a monomer-like conformation over time scales of minutes. According to our mechanism, swapping in GB1 starts from the C-terminus of the polypeptide chain and progresses by exchanging an increasing portion of the chains until a stable conformational state is reached. This exchange process does not involve unfolding. Rather, the conformational changes of individual monomers and their association are tightly coupled to minimize solvent exposure and maximize the total number of native contacts at all times, thereby closely approximating the minimum energy path of the reaction. Using detailed atomic descriptions, we compute the complete free-energy profiles of the exchange reaction for the GB1 quadruple mutant that forms swapped dimers and for the wild-type protein, which is monomeric. In both GB1 forms, intermediates sample a surprisingly wide range of nearly isoenergetic association modes and hinge conformations, indicating that the exchange reaction is a non-specific process akin to encounter complex formation where the amino acid sequence plays a marginal role. The main role of the mutations in the swapping process is to destabilize the GB1 monomer state, while stabilizing the swapped dimer conformation, with non-native intersubunit interactions, fostered by mutant side chains, contributing significantly to this stabilization. Our findings are rationalized in terms of a generic swapping mechanism that involves the association of activated molecular species, and it is argued that a similar mechanism may apply to swapping in other protein systems.

DOI10.1016/j.jmb.2008.06.025
Alternate JournalJ. Mol. Biol.
PubMed ID18588900