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Free keywords:
Binding Sites/genetics
DNA/metabolism
*Evolution, Molecular
Humans
*Hydrophobic and Hydrophilic Interactions
Ligands
Models, Molecular
Multiprotein Complexes/*chemistry/genetics/*metabolism
Mutant Proteins/chemistry/genetics/metabolism
Mutation
Protein Aggregates
Protein Domains
*Protein Multimerization/genetics
Protein Stability
Receptors, Steroid/chemistry/genetics/metabolism
Solvents/chemistry
Abstract:
Most proteins assemble into multisubunit complexes(1). The persistence of these complexes across evolutionary time is usually explained as the result of natural selection for functional properties that depend on multimerization, such as intersubunit allostery or the capacity to do mechanical work(2). In many complexes, however, multimerization does not enable any known function(3). An alternative explanation is that multimers could become entrenched if substitutions accumulate that are neutral in multimers but deleterious in monomers; purifying selection would then prevent reversion to the unassembled form, even if assembly per se does not enhance biological function(3-7). Here we show that a hydrophobic mutational ratchet systematically entrenches molecular complexes. By applying ancestral protein reconstruction and biochemical assays to the evolution of steroid hormone receptors, we show that an ancient hydrophobic interface, conserved for hundreds of millions of years, is entrenched because exposure of this interface to solvent reduces protein stability and causes aggregation, even though the interface makes no detectable contribution to function. Using structural bioinformatics, we show that a universal mutational propensity drives sites that are buried in multimeric interfaces to accumulate hydrophobic substitutions to levels that are not tolerated in monomers. In a database of hundreds of families of multimers, most show signatures of long-term hydrophobic entrenchment. It is therefore likely that many protein complexes persist because a simple ratchet-like mechanism entrenches them across evolutionary time, even when they are functionally gratuitous.
