title: Novel yeast bioassay for high-throughput screening of matrix metalloproteinase inhibitors.
abstract: Diverse malfunctions in the expression and regulation of matrix metalloproteinases (MMPs) are often the cause of severe human diseases, bringing the identification of specific MMP inhibitors into major focus, particularly in anticancer treatment. Here, we describe a novel bioassay based on recombinant yeast cells (Pichia pastoris) that express, deliver, and incorporate biologically active human MMP-2 and MMP-9 at the yeast cell surface. Using Sed1p for cell wall targeting and covalent anchorage, a highly efficient bioassay was established that allows high-throughput screening and subsequent validation of novel MMP inhibitors as potential anticancer drugs. In addition, we developed a straightforward synthesis of a new aspartate-derived MMP inhibitor active in the nM range and bearing an amino functionality that should allow the introduction of a wide range of side chains to modify the properties of these compounds.
authors: Department of Molecular and Cell Biology, Saarland University, P.O. Box 15 11 50, 66041 Saarbruecken, Germany.
authors: Bjoern
authors: B
authors: Diehl
authors: Thorsten M
authors: TM
authors: Hoffmann
authors: Nina C
authors: NC
authors: Mueller
authors: Jens L
authors: JL
authors: Burkhart
authors: Uli
authors: U
authors: Kazmaier
authors: Manfred J
authors: MJ
authors: Schmitt
mesh_terms: D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004791:Enzyme Inhibitors / Q000302:isolation & purification* / Q000494:pharmacology; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D061965:Matrix Metalloproteinase Inhibitors; D020782:Matrix Metalloproteinases / Q000235:genetics; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000235:genetics
chemical_list: D004791:Enzyme Inhibitors; D061965:Matrix Metalloproteinase Inhibitors; D011994:Recombinant Proteins; D020782:Matrix Metalloproteinases