Abstract
Although the development of chemically induced, self-assembled protein-based materials is rapidly expanding, methods for directing their assembly in solution are sparse, and problems of population heterogeneity remain. By exerting control over the assembly of advanced protein structures, new classes of ordered protein nanomaterials become feasible, affecting numerous applications ranging from therapeutics to nanostructural engineering. Focusing on a protein-based method for modulating the stability of a chemically induced dihydrofolate reductase (DHFR) dimer, we demonstrate the sensitivity of a methotrexate competition assay in determining the change in DHFR-DHFR binding cooperativity via interfacial mutations over a 1.3 kcal/mol range. This represents a change of more than 40% of the dimer complex binding energy conferred from protein-protein cooperativity (~3.1 kcal/mol). With the development of this investigative system and refinement of protein-based techniques for complex stability modulation, the directed assembly of protein nanomaterials into heterocomplexes and a concomitant decrease in population heterogeneity becomes a realizable goal.
Copyright © 2012 John Wiley & Sons, Ltd.
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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Amino Acid Substitution / genetics
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Binding, Competitive / drug effects
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Dose-Response Relationship, Drug
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Enzyme Inhibitors / metabolism
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Enzyme Inhibitors / pharmacology
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Escherichia coli Proteins / chemistry
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Escherichia coli Proteins / genetics
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Escherichia coli Proteins / metabolism
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Folic Acid Antagonists / metabolism
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Folic Acid Antagonists / pharmacology
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Methotrexate / metabolism
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Methotrexate / pharmacology*
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Models, Molecular
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Mutagenesis, Site-Directed
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Mutant Proteins / chemistry
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Mutant Proteins / metabolism
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Osmolar Concentration
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Protein Interaction Domains and Motifs / drug effects*
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Protein Interaction Domains and Motifs / genetics
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Protein Interaction Domains and Motifs / physiology
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Protein Multimerization / drug effects*
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Protein Multimerization / genetics
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Protein Structure, Quaternary / drug effects
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Tetrahydrofolate Dehydrogenase / chemistry
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Tetrahydrofolate Dehydrogenase / drug effects
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Tetrahydrofolate Dehydrogenase / genetics
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Tetrahydrofolate Dehydrogenase / metabolism*
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Up-Regulation / drug effects
Substances
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Enzyme Inhibitors
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Escherichia coli Proteins
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Folic Acid Antagonists
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Mutant Proteins
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Tetrahydrofolate Dehydrogenase
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Methotrexate