A systematic method for identifying small-molecule modulators of protein–protein interactions
AUTOR(ES)
Horswill, Alexander R.
FONTE
National Academy of Sciences
RESUMO
Discovering small-molecule modulators of protein–protein interactions is a challenging task because of both the generally noncontiguous, large protein surfaces that form these interfaces and the shortage of high-throughput approaches capable of identifying such rare inhibitors. We describe here a robust and flexible methodology that couples disruption of protein–protein complexes to host cell survival. The feasibility of this approach was demonstrated through monitoring a small-molecule-mediated protein–protein association (FKBP12–rapamycin–FRAP) and two cases of dissociation (homodimeric HIV-1 protease and heterodimeric ribonucleotide reductase). For ribonucleotide reductase, we identified cyclic peptide inhibitors from genetically encoded libraries that dissociated the enzyme subunits. A solid-phase synthetic strategy and peptide ELISAs were developed to characterize these inhibitors, resulting in the discovery of cyclic peptides that operate in an unprecedented manner, thus highlighting the strengths of a functional approach. The ability of this method to process large libraries, coupled with the benefits of a genetic selection, allowed us to identify rare, uniquely active small-molecule modulators of protein–protein interactions at a frequency of less than one in 10 million.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=524857Documentos Relacionados
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