Mechanistic and Structural Features of PROTAC Ternary Complexes

Ryan Casement, Adam Bond, Conner Craigon, Alessio Ciulli (Lead / Corresponding author)

Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)peer-review

Abstract

The rapid and ever-growing advancements from within the field of proteolysis-targeting chimeras (PROTAC)-induced protein degradation have driven considerable development to gain a deeper understanding of their mode of action. The ternary complex formed by PROTACs with their target protein and E3 ubiquitin ligase is the key species in their substoichiometric catalytic mechanism. Here, we describe the theoretical framework that underpins ternary complexes, including a current understanding of the three-component binding model, cooperativity, hook effect and structural considerations. We discuss in detail the biophysical methods used to interrogate ternary complex formation in vitro, including X-ray crystallography, AlphaLISA, FRET, FP, ITC and SPR. Finally, we provide detailed ITC methods and discuss approaches to assess binary and ternary target engagement, target ubiquitination and degradation that can be used to obtain a more holistic understanding of the mode of action within a cellular environment.

Original languageEnglish
Title of host publicationTargeted Protein Degradation
Subtitle of host publicationMethods and Protocols
EditorsAngela M. Cacace, Christopher M. Hickey, Miklós Békés
Place of PublicationNew York
PublisherHumana Press
Pages79-113
Number of pages35
Edition1
ISBN (Electronic)9781071616659
ISBN (Print)9781071616642 (hbk), 9781071616673 (pbk)
DOIs
Publication statusPublished - 2021

Publication series

NameMethods in Molecular Biology
Volume2365
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Biophysical methods, crystal structures
  • Cooperativity
  • E3 ubiquitin ligase
  • Hook effect
  • Protein ubiquitination
  • Target engagement
  • Targeted protein degradation (TPD)
  • Ternary complex
  • proteolysis-targeting chimeras (PROTACs)

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