title: Reversible Inhibitors Arrest ClpP in a Defined Conformational State that Can Be Revoked by ClpX Association.
abstract: Caseinolytic protease P (ClpP) is an important regulator of Staphylococcus aureus pathogenesis. A high-throughput screening for inhibitors of ClpP peptidase activity led to the identification of the first non-covalent binder for this enzyme class. Co-crystallization of the small molecule with S. aureus ClpP revealed a novel binding mode: Because of the rotation of the conserved residue proline 125, ClpP is locked in a defined conformational state, which results in distortion of the catalytic triad and inhibition of the peptidase activity. Based on these structural insights, the molecule was optimized by rational design and virtual screening, resulting in derivatives exceeding the potency of previous ClpP inhibitors. Strikingly, the conformational lock is overturned by binding of ClpX, an associated chaperone that enables proteolysis by substrate unfolding in the ClpXP complex. Thus, regulation of inhibitor binding by associated chaperones is an unexpected mechanism important for ClpP drug development.
authors.affiliation: Center for Integrated Protein Science at the Department of Chemistry, Technische Universität München, Lichtenbergstrasse 4, 85747 Garching (Germany).
authors.forename: Axel
authors.initials: A
authors.lastname: Pahl
authors.affiliation: Center for Integrated Protein Science at the Department of Chemistry, Technische Universität München, Lichtenbergstrasse 4, 85747 Garching (Germany).
authors.forename: Markus
authors.initials: M
authors.lastname: Lakemeyer
authors.affiliation: Center for Integrated Protein Science at the Department of Chemistry, Technische Universität München, Lichtenbergstrasse 4, 85747 Garching (Germany).
authors.forename: Marie-Theres
authors.initials: MT
authors.lastname: Vielberg
authors.affiliation: Center for Integrated Protein Science at the Department of Chemistry, Technische Universität München, Lichtenbergstrasse 4, 85747 Garching (Germany).
authors.forename: Mathias W
authors.initials: MW
authors.lastname: Hackl
authors.affiliation: Center for Integrated Protein Science at the Department of Chemistry, Technische Universität München, Lichtenbergstrasse 4, 85747 Garching (Germany).
authors.forename: Jan
authors.initials: J
authors.lastname: Vomacka
authors.affiliation: Center for Integrated Protein Science at the Department of Chemistry, Technische Universität München, Lichtenbergstrasse 4, 85747 Garching (Germany).
authors.forename: Vadim S
authors.initials: VS
authors.lastname: Korotkov
authors.affiliation: Center for Integrated Protein Science at the Department of Chemistry, Technische Universität München, Lichtenbergstrasse 4, 85747 Garching (Germany).
authors.forename: Martin L
authors.initials: ML
authors.lastname: Stein
authors.affiliation: Center for Integrated Protein Science at the Department of Chemistry, Technische Universität München, Lichtenbergstrasse 4, 85747 Garching (Germany).
authors.forename: Christian
authors.initials: C
authors.lastname: Fetzer
authors.affiliation: Center for Integrated Protein Science at the Department of Chemistry, Technische Universität München, Lichtenbergstrasse 4, 85747 Garching (Germany).
authors.forename: Katrin
authors.initials: K
authors.lastname: Lorenz-Baath
authors.affiliation: Center for Integrated Protein Science at the Department of Chemistry, Technische Universität München, Lichtenbergstrasse 4, 85747 Garching (Germany).
authors.forename: Klaus
authors.initials: K
authors.lastname: Richter
authors.affiliation: Department of Chemistry and Chemical Biology, Technische Universität Dortmund, Otto-Hahn-Strasse 6, 44221 Dortmund (Germany).
authors.forename: Herbert
authors.initials: H
authors.lastname: Waldmann
authors.affiliation: Center for Integrated Protein Science at the Department of Chemistry, Technische Universität München, Lichtenbergstrasse 4, 85747 Garching (Germany). michael.groll@tum.de.
authors.forename: Michael
authors.initials: M
authors.lastname: Groll
authors.affiliation: Center for Integrated Protein Science at the Department of Chemistry, Technische Universität München, Lichtenbergstrasse 4, 85747 Garching (Germany). stephan.sieber@tum.de.
authors.forename: Stephan A
authors.initials: SA
authors.lastname: Sieber
mesh_terms: D011487:Protein Conformation; D012697:Serine Endopeptidases / Q000187:drug effects*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology*; D013329:Structure-Activity Relationship
chemical_list: D015842:Serine Proteinase Inhibitors; D012697:Serine Endopeptidases
keywords: caseinolytic protease; conformational selection; non-covalent inhibition; protein crystallography; structural rearrangement