iBRET Screen of the ABCD1 Peroxisomal Network and Mutation-Induced Network Perturbations

Amelie S. Lotz-Havla, Mathias Woidy, Philipp Guder, Caroline C. Friedel, Julian M. Klingbeil, Ana Maria Bulau, Anja Schultze, Ilona Dahmen, Heidi Noll-Puchta, Stephan Kemp, Ralf Erdmann, Ralf Zimmer, Ania C. Muntau, Søren W. Gersting

Research output: Contribution to JournalArticleAcademicpeer-review

3 Citations (Scopus)

Abstract

Mapping the network of proteins provides a powerful means to investigate the function of disease genes and to unravel the molecular basis of phenotypes. We present an automated informatics-aided and bioluminescence resonance energy transfer-based approach (iBRET) enabling high-confidence detection of protein-protein interactions in living mammalian cells. A screen of the ABCD1 protein, which is affected in X-linked adrenoleukodystrophy (X-ALD), against an organelle library of peroxisomal proteins demonstrated applicability of iBRET for large-scale experiments. We identified novel protein-protein interactions for ABCD1 (with ALDH3A2, DAO, ECI2, FAR1, PEX10, PEX13, PEX5, PXMP2, and PIPOX), mapped its position within the peroxisomal protein-protein interaction network, and determined that pathogenic missense variants in ABCD1 alter the interaction with selected binding partners. These findings provide mechanistic insights into pathophysiology of X-ALD and may foster the identification of new disease modifiers.

Original languageEnglish
Pages (from-to)4366-4380
Number of pages15
JournalJournal of Proteome Research
Volume20
Issue number9
DOIs
Publication statusPublished - 3 Sep 2021

Keywords

  • ABCD1
  • BRET
  • FAR1
  • X-ALD
  • bioluminescence resonance energy transfer
  • fatty acids
  • interactome
  • lipid droplets
  • living cells
  • protein-protein interaction
  • screening

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