Designing protein dimerizers: The importance of ligand conformational equilibria

Jonathan C.T. Carlson, Aaron Kanter, Guruvasuthevan R. Thuduppathy, Vivian Cody, Pamela E. Pineda, R S Mc Ivor, Carston R Wagner

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

In an effort to elucidate the role of ligand conformation in induced protein dimerization, we synthesized a flexible methotrexate (MTX) dimer, demonstrated its ability to selectively dimerize Escherichia coli dihydrofolate reductase (DHFR), and evaluated the factors regulating its ability to induce cooperative dimerization. Despite known entropic barriers, bis-MTX proved to possess substantial conformational stability in aqueous solution (-3.8 kcal/mol ≥ ΔGfold ≥ -4.9 kcal/mol), exerting a dominant influence on the thermodynamics of dimerization. To dimerize DHFR, bis-MTX must shift from a folded to an extended conformation. From this conclusion, the strength of favorable protein-protein interactions in bis-MTX-E. coli DHFR dimers (-3.1 kcal/mol ≥ ΔGc ≥ -4.2 kcal/mol), and the selectivity of dimerization for E. coli DHFR relative to mouse DHFR (>107) could be determined. The crystal structure of bis-MTX in complex with E. coli DHFR confirms the feasibility of a close-packed dimerization interface and suggests a possible solution conformation for the induced protein dimers. Consequently, the secondary structure of this minimal foldamer regulates its ability to dimerize dihydrofolate reductase in solution, providing insight into the complex energy landscape of induced dimerization.

Original languageEnglish (US)
Pages (from-to)1501-1507
Number of pages7
JournalJournal of the American Chemical Society
Volume125
Issue number6
DOIs
StatePublished - Feb 12 2003

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Tetrahydrofolate Dehydrogenase
Dimerization
Ligands
Methotrexate
Proteins
Escherichia coli
Dimers
Conformations
Protein Multimerization
Protein Conformation
Oxidoreductases
Thermodynamics
Crystal structure

Cite this

Designing protein dimerizers : The importance of ligand conformational equilibria. / Carlson, Jonathan C.T.; Kanter, Aaron; Thuduppathy, Guruvasuthevan R.; Cody, Vivian; Pineda, Pamela E.; Mc Ivor, R S; Wagner, Carston R.

In: Journal of the American Chemical Society, Vol. 125, No. 6, 12.02.2003, p. 1501-1507.

Research output: Contribution to journalArticle

Carlson, Jonathan C.T. ; Kanter, Aaron ; Thuduppathy, Guruvasuthevan R. ; Cody, Vivian ; Pineda, Pamela E. ; Mc Ivor, R S ; Wagner, Carston R. / Designing protein dimerizers : The importance of ligand conformational equilibria. In: Journal of the American Chemical Society. 2003 ; Vol. 125, No. 6. pp. 1501-1507.
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