Discovery of antagonist peptides against bacterial helicase-primase interaction in B. stearothermophilus by reverse yeast three-hybrid.
Developing small-molecule antagonists against protein-protein interactions will provide powerful tools for mechanistic/functional studies and the discovery of new antibacterials. We have developed a reverse yeast three-hybrid approach that allows high-throughput screening for antagonist peptides against essential protein-protein interactions. We have applied our methodology to the essential bacterial helicase-primase interaction in Bacillus stearothermophilus and isolated a unique antagonist peptide. This peptide binds to the primase, thus excluding the helicase and inhibiting an essential interaction in bacterial DNA replication. We provide proof of principle that our reverse yeast three-hybrid method is a powerful "one-step" screen tool for direct high-throughput antagonist peptide selection against any protein-protein interaction detectable by traditional yeast two-hybrid systems. Such peptides will provide useful "leads" for the development of new antibacterials.
Gardiner
Laurence
L
School of Chemistry, Centre for Biomolecular Sciences, University of Nottingham, University Park, Nottingham NG7 2RD, United Kingdom.
Coyle
Barry J
BJ
Chan
Weng C
WC
Soultanas
Panos
P
D000251:Adenosine Triphosphatases / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism; D001667:Binding, Competitive; D004265:DNA Helicases / Q000378:metabolism*; D019915:DNA Primase / Q000378:metabolism*; D001411:Geobacillus stearothermophilus / Q000201:enzymology; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000378:metabolism*; D019151:Peptide Library; D010455:Peptides / Q000138:chemical synthesis / Q000378:metabolism*; D011485:Protein Binding; D012441:Saccharomyces cerevisiae; D020798:Two-Hybrid System Techniques
D001426:Bacterial Proteins; D010446:Peptide Fragments; D019151:Peptide Library; D010455:Peptides; D019915:DNA Primase; D000251:Adenosine Triphosphatases; D004265:DNA Helicases