Native mass spectrometry of complexes formed by molecular glues reveals stoichiometric rearrangement of E3 ligases

Jackson, Cara and Beveridge, Rebecca (2024) Native mass spectrometry of complexes formed by molecular glues reveals stoichiometric rearrangement of E3 ligases. Analyst, 149 (11). pp. 3178-3185. ISSN 0003-2654 (https://doi.org/10.1039/d4an00110a)

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Abstract

In this application of native mass spectrometry (nMS) to investigate complexes formed by molecular glues (MGs), we have demonstrated its efficiency in delineating stoichiometric rearrangements of E3 ligases that occur during targeted protein degradation (TPD). MGs stabilise interactions between an E3 ligase and a protein of interest (POI) targeted for degradation, and these ternary interactions are challenging to characterise. We have shown that nMS can unambiguously identify complexes formed between the CRBN : DDB1 E3 ligase and the POI GSPT1 upon the addition of lenalidomide, pomalidomide or thalidomide. Ternary complex formation was also identified involving the DCAF15 : DDA1 : DDB1 E3 ligase in the presence of MG (E7820 or indisulam) and POI RBM39. Moreover, we uncovered that the DCAF15 : DDA1 : DDB1 E3 ligase self-associates into dimers and trimers when analysed alone at low salt concentrations (100 mM ammonium acetate) which dissociate into single copies of the complex at higher salt concentrations (500 mM ammonium acetate), or upon the addition of MG and POI, forming a 1 : 1 : 1 ternary complex. This work demonstrates the strength of nMS in TPD research, reveals novel binding mechanisms of the DCAF15 E3 ligase, and its self-association into dimers and trimers at reduced salt concentration during structural analysis.

ORCID iDs

Beveridge, Rebecca ORCID: https://orcid.org/0000-0003-0320-6496

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• Item metadata

  Item type:	Article
  ID code:	88894
  Dates:	Date Event 16 April 2024 Published 16 April 2024 Published Online 15 April 2024 Accepted 22 January 2024 Submitted
  Subjects:	Science > Chemistry
  Department:	Faculty of Science > Pure and Applied Chemistry
  Depositing user:	Pure Administrator
  Date deposited:	22 Apr 2024 14:30
  Last modified:	11 Nov 2024 14:17
  URI:	https://strathprints.strath.ac.uk/id/eprint/88894