title: Binding mode of TMC-95A analogues to eukaryotic 20S proteasome.
abstract: The complex thermodynamics that govern noncovalent protein-ligand interactions are still not fully understood, despite the exponential increase in experimental structural data available from X-ray crystallography and NMR spectroscopy. The eukaryotic 20S proteasome offers an ideal system for such studies as it contains in duplicate three proteolytically active sites with different substrate specificities. The natural product TMC-95A inhibits these proteolytic centers noncovalently with distinct affinities. X-ray crystallographic analysis of the complexes of the yeast proteasome core particle with this natural inhibitor and two synthetic analogues clearly revealed highly homologous hydrogen-bonding networks involving mainly the peptide backbone despite the strongly differentiated binding affinities to the three active sites of the 20S proteasome. The natural product and the two analogues are constrained in a rigid beta-type extended conformation by the endocyclic biaryl clamp, which preorganizes the peptide backbone for optimal adaptation of the ligands to the active site clefts and thus favors the binding processes entropically. However, the biaryl clamp also dictates the orientation of the P1 and P3 residues and their mode of interaction with the protein binding subsites. This limitation is optimally solved in TMC-95A with the conformationally restricted (Z)-prop-1-enyl group acting as P1 residue, at least for the chymotrypsin-like active site; however, it critically affects the inhibitory potencies of the analogues, thus suggesting the use of less-rigid endocyclic clamps in the design of proteasome inhibitors that allow for a better presentation of residues interacting with the active site clefts of the enzyme.
authors.affiliation: Max-Planck-Institut für Biochemie, Am Klopferspitz 18, 82152 Martinsried, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Kaiser
authors.forename: Michael
authors.initials: M
authors.lastname: Groll
authors.forename: Carlo
authors.initials: C
authors.lastname: Siciliano
authors.forename: Irmgard
authors.initials: I
authors.lastname: Assfalg-Machleidt
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Weyher
authors.forename: Jun
authors.initials: J
authors.lastname: Kohno
authors.forename: Alexander G
authors.initials: AG
authors.lastname: Milbradt
authors.forename: Christian
authors.initials: C
authors.lastname: Renner
authors.forename: Robert
authors.initials: R
authors.lastname: Huber
authors.forename: Luis
authors.initials: L
authors.lastname: Moroder
mesh_terms: D001665:Binding Sites; D018360:Crystallography, X-Ray; D015195:Drug Design; D005057:Eukaryotic Cells / Q000378:metabolism; D007202:Indicators and Reagents; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D010456:Peptides, Cyclic / Q000378:metabolism* / Q000494:pharmacology; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D061988:Proteasome Inhibitors; D011485:Protein Binding; D015003:Yeasts / Q000378:metabolism
chemical_list: D007202:Indicators and Reagents; D010456:Peptides, Cyclic; D061988:Proteasome Inhibitors; C408291:TMC-95A; D046988:Proteasome Endopeptidase Complex