5AFG

Structure of the Stapled Peptide Bound to Mdm2

Summary for 5AFG

• Entry DOI: 10.2210/pdb5afg/pdb
• Descriptor: E3 UBIQUITIN-PROTEIN LIGASE MDM2, STAPLED PEPTIDE, 1,8-DIETHYL-1,8-DIHYDRODIBENZO[3,4:7,8][1,2,3]TRIAZOLO[4',5':5,6]CYCLOOCTA[1,2-D][1,2,3]TRIAZOLE, ... (4 entities in total)
• Functional Keywords: ligase, mdm2
• Biological source: HOMO SAPIENS (HUMAN); More
• Total number of polymer chains: 2
• Total formula weight: 12865.93

Authors

Lau, Y.H.,Wu, Y.,Rossmann, M.,de Andrade, P.,Tan, Y.S.,McKenzie, G.J.,Venkitaraman, A.R.,Hyvonen, M.,Spring, D.R. (deposition date: 2015-01-22, release date: 2016-01-27, Last modification date: 2024-01-10)

Primary citation

Lau, Y.H.,Wu, Y.,Rossmann, M.,Tan, B.X.,De Andrade, P.,Tan, Y.S.,Verma, C.,Mckenzie, G.J.,Venkitaraman, A.R.,Hyvonen, M.,Spring, D.R.
Double Strain-Promoted Macrocyclization for the Rapid Selection of Cell-Active Stapled Peptides.
Angew.Chem.Int.Ed.Engl., 54:15410-, 2015

PubMed Abstract: Peptide stapling is a method for designing macrocyclic alpha-helical inhibitors of protein-protein interactions. However, obtaining a cell-active inhibitor can require significant optimization. We report a novel stapling technique based on a double strain-promoted azide-alkyne reaction, and exploit its biocompatibility to accelerate the discovery of cell-active stapled peptides. As a proof of concept, MDM2-binding peptides were stapled in parallel, directly in cell culture medium in 96-well plates, and simultaneously evaluated in a p53 reporter assay. This in situ stapling/screening process gave an optimal candidate that showed improved proteolytic stability and nanomolar binding to MDM2 in subsequent biophysical assays. α-Helicity was confirmed by a crystal structure of the MDM2-peptide complex. This work introduces in situ stapling as a versatile biocompatible technique with many other potential high-throughput biological applications.

PubMed: 26768531
DOI: 10.1002/ANIE.201508416

Experimental method

X-RAY DIFFRACTION (1.9 Å)