title: Fragment-based screening using surface plasmon resonance technology.
abstract: Surface plasmon resonance (SPR) technology has emerged as a new and powerful technique to investigate the interaction between low-molecular-weight molecules and target proteins. In the present work, the authors assemble from a large compound collection a library of 2226 molecules (fragments having low molecular weights between 100 and 300 Da) to screen them for binding to chymase, a serine protease. Both the active chymase and a zymogen-like form of the protein were used in parallel to distinguish between specific and unspecific binding. The relative ligand-binding activity of the immobilized protein was periodically measured with a reference compound. The screening experiments were performed at 25 degrees C at a fragment concentration of 200 microM in the presence of 2% DMSO. Applying the filter cascade, affinity-selectivity-competition (competition with reference compounds and cross-competition with fragments), 80 compounds show up as positive screening hits. Competition experiments between fragments show that they bind to different parts of the active site. Of 36 fragments co-crystallized for X-ray studies, 12 could be located in the active site of the protein. These results validate the authors' library and demonstrate that the application of SPR technology as a filter in fragment screening can be achieved successfully.
authors.affiliation: F. Hoffmann-La Roche Ltd, Pharma Research, Discovery Technologies, Basel, Switzerland. samantha.perspicace@unibas.ch
authors.forename: Samantha
authors.initials: S
authors.lastname: Perspicace
authors.forename: David
authors.initials: D
authors.lastname: Banner
authors.forename: Jörg
authors.initials: J
authors.lastname: Benz
authors.forename: Francis
authors.initials: F
authors.lastname: Müller
authors.forename: Daniel
authors.initials: D
authors.lastname: Schlatter
authors.forename: Walter
authors.initials: W
authors.lastname: Huber
mesh_terms: D000327:Adsorption; D053818:Chymases / Q000378:metabolism; D018360:Crystallography, X-Ray; D005453:Fluorescence; D006801:Humans; D008024:Ligands; D008970:Molecular Weight; D011485:Protein Binding; D012015:Reference Standards; D015203:Reproducibility of Results; D054852:Small Molecule Libraries / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D020349:Surface Plasmon Resonance / Q000379:methods*; D013696:Temperature; D017186:Titrimetry
chemical_list: D008024:Ligands; D054852:Small Molecule Libraries; D053818:Chymases