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Research output: Contribution to Journal/Magazine › Journal article › peer-review
Research output: Contribution to Journal/Magazine › Journal article › peer-review
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TY - JOUR
T1 - Structural basis of PROTAC cooperative recognition for selective protein degradation
AU - Gadd, Morgan S
AU - Testa, Andrea
AU - Lucas, Xavier
AU - Chan, Kwok-Ho
AU - Chen, Wenzhang
AU - Lamont, Douglas J
AU - Zengerle, Michael
AU - Ciulli, Alessio
PY - 2017/5/31
Y1 - 2017/5/31
N2 - Inducing macromolecular interactions with small molecules to activate cellular signaling is a challenging goal. PROTACs (proteolysis-targeting chimeras) are bifunctional molecules that recruit a target protein in proximity to an E3 ubiquitin ligase to trigger protein degradation. Structural elucidation of the key ternary ligase-PROTAC-target species and its impact on target degradation selectivity remain elusive. We solved the crystal structure of Brd4 degrader MZ1 in complex with human VHL and the Brd4 bromodomain (Brd4BD2). The ligand folds into itself to allow formation of specific intermolecular interactions in the ternary complex. Isothermal titration calorimetry studies, supported by surface mutagenesis and proximity assays, are consistent with pronounced cooperative formation of ternary complexes with Brd4BD2. Structure-based-designed compound AT1 exhibits highly selective depletion of Brd4 in cells. Our results elucidate how PROTAC-induced de novo contacts dictate preferential recruitment of a target protein into a stable and cooperative complex with an E3 ligase for selective degradation.
AB - Inducing macromolecular interactions with small molecules to activate cellular signaling is a challenging goal. PROTACs (proteolysis-targeting chimeras) are bifunctional molecules that recruit a target protein in proximity to an E3 ubiquitin ligase to trigger protein degradation. Structural elucidation of the key ternary ligase-PROTAC-target species and its impact on target degradation selectivity remain elusive. We solved the crystal structure of Brd4 degrader MZ1 in complex with human VHL and the Brd4 bromodomain (Brd4BD2). The ligand folds into itself to allow formation of specific intermolecular interactions in the ternary complex. Isothermal titration calorimetry studies, supported by surface mutagenesis and proximity assays, are consistent with pronounced cooperative formation of ternary complexes with Brd4BD2. Structure-based-designed compound AT1 exhibits highly selective depletion of Brd4 in cells. Our results elucidate how PROTAC-induced de novo contacts dictate preferential recruitment of a target protein into a stable and cooperative complex with an E3 ligase for selective degradation.
KW - Amino Acid Sequence
KW - Crystallography, X-Ray
KW - Dipeptides
KW - Elongin
KW - Heterocyclic Compounds, 3-Ring
KW - Humans
KW - Models, Molecular
KW - Multiprotein Complexes
KW - Nuclear Proteins
KW - Protein Binding
KW - Protein Conformation
KW - Proteolysis
KW - Small Molecule Libraries
KW - Structure-Activity Relationship
KW - Thermodynamics
KW - Transcription Factors
KW - Ubiquitin-Protein Ligases
KW - Von Hippel-Lindau Tumor Suppressor Protein
U2 - 10.1038/nchembio.2329
DO - 10.1038/nchembio.2329
M3 - Journal article
C2 - 28288108
VL - 13
SP - 514
EP - 521
JO - Nature Chemical Biology
JF - Nature Chemical Biology
SN - 1552-4450
IS - 5
ER -