TY - JOUR
T1 - iBRET Screen of the ABCD1 Peroxisomal Network and Mutation-Induced Network Perturbations
AU - Lotz-Havla, Amelie S.
AU - Woidy, Mathias
AU - Guder, Philipp
AU - Friedel, Caroline C.
AU - Klingbeil, Julian M.
AU - Bulau, Ana Maria
AU - Schultze, Anja
AU - Dahmen, Ilona
AU - Noll-Puchta, Heidi
AU - Kemp, Stephan
AU - Erdmann, Ralf
AU - Zimmer, Ralf
AU - Muntau, Ania C.
AU - Gersting, Søren W.
N1 - Funding Information: This work was supported by the Bavarian Genome Research Network (BayGene) and an LMUexcellent grant 42595-6 to A.C.M., by a Prinz Lennart von Hohenzollern Stiftung grant to A.C.M and S.W.G. as well as a Fritz Thyssen Stiftung scholarship to A.S.L.-H. Publisher Copyright: © 2021 American Chemical Society. Copyright: Copyright 2021 Elsevier B.V., All rights reserved.
PY - 2021/9/3
Y1 - 2021/9/3
N2 - 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.
AB - 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.
KW - ABCD1
KW - BRET
KW - FAR1
KW - X-ALD
KW - bioluminescence resonance energy transfer
KW - fatty acids
KW - interactome
KW - lipid droplets
KW - living cells
KW - protein-protein interaction
KW - screening
UR - http://www.scopus.com/inward/record.url?scp=85113957417&partnerID=8YFLogxK
U2 - https://doi.org/10.1021/acs.jproteome.1c00330
DO - https://doi.org/10.1021/acs.jproteome.1c00330
M3 - Article
C2 - 34383492
SN - 1535-3893
VL - 20
SP - 4366
EP - 4380
JO - Journal of proteome research
JF - Journal of proteome research
IS - 9
ER -