title: High-Throughput Screening (HTS) by NMR Guided Identification of Novel Agents Targeting the Protein Docking Domain of YopH.
abstract: Recently we described a novel approach, named high-throughput screening (HTS) by NMR that allows the identification, from large combinatorial peptide libraries, of potent and selective peptide mimetics against a given target. Here, we deployed the "HTS by NMR" approach for the design of novel peptoid sequences targeting the N-terminal domain of Yersinia outer protein H (YopH-NT), a bacterial toxin essential for the virulence of Yersinia pestis. We aimed at disrupting the protein-protein interactions between YopH-NT and its cellular substrates, with the goal of inhibiting indirectly YopH enzymatic function. These studies resulted in a novel agent of sequence Ac-F-pY-cPG-d-P-NH2 (pY=phosphotyrosine; cPG=cyclopentyl glycine) with a Kd value against YopH-NT of 310 nm. We demonstrated that such a pharmacological inhibitor of YopH-NT results in the inhibition of the dephosphorylation by full-length YopH of a cellular substrate. Hence, potentially this agent represents a valuable stepping stone for the development of novel therapeutics against Yersinia infections. The data reported further demonstrate the utility of the HTS by NMR approach in deriving novel peptide mimetics targeting protein-protein interactions.
authors.affiliation: Sanford Burnham Prebys Medical Discovery Institute, 10901 North Torrey Pines Road, La Jolla, CA, 92037, USA.|Sanford Burnham Prebys Graduate School of Biomedical Sciences, 10901 North Torrey Pines Road, La Jolla, CA, 92037, USA.
authors.forename: Angel
authors.initials: A
authors.lastname: Bottini
authors.affiliation: Sanford Burnham Prebys Medical Discovery Institute, 10901 North Torrey Pines Road, La Jolla, CA, 92037, USA.
authors.forename: Bainan
authors.initials: B
authors.lastname: Wu
authors.affiliation: Sanford Burnham Prebys Medical Discovery Institute, 10901 North Torrey Pines Road, La Jolla, CA, 92037, USA.|Division of Biomedical Sciences, School of Medicine, University of California Riverside, 900 University Avenue, Riverside, CA, 92521, USA.
authors.forename: Elisa
authors.initials: E
authors.lastname: Barile
authors.affiliation: Sanford Burnham Prebys Medical Discovery Institute, 10901 North Torrey Pines Road, La Jolla, CA, 92037, USA.|Division of Biomedical Sciences, School of Medicine, University of California Riverside, 900 University Avenue, Riverside, CA, 92521, USA.
authors.forename: Surya K
authors.initials: SK
authors.lastname: De
authors.affiliation: Institute of Biostructures & Bioimaging, National Research Council (IBB-CNR), Via De Amicis 95, Naples, 80145, Italy.
authors.forename: Marilisa
authors.initials: M
authors.lastname: Leone
authors.affiliation: Sanford Burnham Prebys Medical Discovery Institute, 10901 North Torrey Pines Road, La Jolla, CA, 92037, USA. maurizio.pellecchia@ucr.edu.|Division of Biomedical Sciences, School of Medicine, University of California Riverside, 900 University Avenue, Riverside, CA, 92521, USA. maurizio.pellecchia@ucr.edu.
authors.forename: Maurizio
authors.initials: M
authors.lastname: Pellecchia
mesh_terms: D000900:Anti-Bacterial Agents / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D057166:High-Throughput Screening Assays; D009682:Magnetic Resonance Spectroscopy; D008826:Microbial Sensitivity Tests; D019906:Nuclear Magnetic Resonance, Biomolecular; D034444:Peptoids / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D010930:Plague / Q000188:drug therapy; D011485:Protein Binding / Q000187:drug effects; D000072417:Protein Domains / Q000187:drug effects; D017027:Protein Tyrosine Phosphatases / Q000737:chemistry* / Q000378:metabolism*; D013329:Structure-Activity Relationship; D015010:Yersinia pestis / Q000187:drug effects
chemical_list: D000900:Anti-Bacterial Agents; D001425:Bacterial Outer Membrane Proteins; D034444:Peptoids; D017027:Protein Tyrosine Phosphatases; C067571:yopH protein, Yersinia
keywords: NMR spectroscopy; YopH; combinatorial libraries; high-throughput screening (HTS); peptides; protein-protein interactions