title: Cullin3-BTB interface: a novel target for stapled peptides.
abstract: Cullin3 (Cul3), a key factor of protein ubiquitination, is able to interact with dozens of different proteins containing a BTB (Bric-a-brac, Tramtrack and Broad Complex) domain. We here targeted the Cul3-BTB interface by using the intriguing approach of stabilizing the α-helical conformation of Cul3-based peptides through the "stapling" with a hydrocarbon cross-linker. In particular, by combining theoretical and experimental techniques, we designed and characterized stapled Cul3-based peptides embedding the helix 2 of the protein (residues 49-68). Intriguingly, CD and NMR experiments demonstrate that these stapled peptides were able to adopt the helical structure that the fragment assumes in the parent protein. We also show that some of these peptides were able to bind to the BTB of the tetrameric KCTD11, a substrate adaptor involved in HDAC1 degradation, with high affinity (~ 300-600 nM). Cul3-derived staple peptides are also able to bind the BTB of the pentameric KCTD5. Interestingly, the affinity of these peptides is of the same order of magnitude of that reported for the interaction of full-length Cul3 with some BTB containing proteins. Moreover, present data indicate that stapling endows these peptides with an increased serum stability. Altogether, these findings indicate that the designed stapled peptides can efficiently mimic protein-protein interactions and are potentially able to modulate fundamental biological processes involving Cul3.
authors.affiliation: Institute of Biostructures and Bioimaging, C.N.R., Napoli, Italy.
authors.forename: Ivan
authors.initials: I
authors.lastname: de Paola
authors.affiliation: Institute of Crystallography, C.N.R., Bari, Italy.
authors.forename: Luciano
authors.initials: L
authors.lastname: Pirone
authors.affiliation: Second University of Napoli, Caserta, Italy.
authors.forename: Maddalena
authors.initials: M
authors.lastname: Palmieri
authors.affiliation: Institute of Biostructures and Bioimaging, C.N.R., Napoli, Italy; Second University of Napoli, Caserta, Italy.
authors.forename: Nicole
authors.initials: N
authors.lastname: Balasco
authors.affiliation: Institute of Biostructures and Bioimaging, C.N.R., Napoli, Italy; Interuniversity Centre for Research on Bioactive Peptides (CIRPEB), Napoli, Italy.
authors.forename: Luciana
authors.initials: L
authors.lastname: Esposito
authors.affiliation: Second University of Napoli, Caserta, Italy.
authors.forename: Luigi
authors.initials: L
authors.lastname: Russo
authors.affiliation: Department of Molecular Medicine, La Sapienza University, Roma, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Mazzà
authors.affiliation: Department of Molecular Medicine, La Sapienza University, Roma, Italy.
authors.forename: Lucia
authors.initials: L
authors.lastname: Di Marcotullio
authors.affiliation: Institute of Biostructures and Bioimaging, C.N.R., Napoli, Italy; Interuniversity Centre for Research on Bioactive Peptides (CIRPEB), Napoli, Italy.
authors.forename: Sonia
authors.initials: S
authors.lastname: Di Gaetano
authors.affiliation: Second University of Napoli, Caserta, Italy.
authors.forename: Gaetano
authors.initials: G
authors.lastname: Malgieri
authors.affiliation: Institute of Biostructures and Bioimaging, C.N.R., Napoli, Italy; Interuniversity Centre for Research on Bioactive Peptides (CIRPEB), Napoli, Italy.
authors.forename: Luigi
authors.initials: L
authors.lastname: Vitagliano
authors.affiliation: Institute of Biostructures and Bioimaging, C.N.R., Napoli, Italy; Interuniversity Centre for Research on Bioactive Peptides (CIRPEB), Napoli, Italy.
authors.forename: Emilia
authors.initials: E
authors.lastname: Pedone
authors.affiliation: Institute of Biostructures and Bioimaging, C.N.R., Napoli, Italy; Interuniversity Centre for Research on Bioactive Peptides (CIRPEB), Napoli, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Zaccaro
mesh_terms: D044842:Cullin Proteins / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary
chemical_list: C503645:CUL3 protein, human; D044842:Cullin Proteins; D010455:Peptides
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