TY - JOUR
T1 - Identification of Full-Length Wild-Type and Mutant Huntingtin Interacting Proteins by Crosslinking Immunoprecipitation in Mice Brain Cortex
AU - Sap, Karen A.
AU - Guler, Arzu Tugce
AU - Bury, Aleksandra
AU - Dekkers, Dick
AU - Demmers, Jeroen A. A.
AU - Reits, Eric A.
N1 - Funding Information: In this work we reported both shared and unique wtHtt and mHtt interactors, which are involved in various cellular processes, including exocytosis, transport of vesicles, translation and metabolism Figure 1 and Fig. 4 were created with BioRen-der.com. Figure 2C was created with VIB-UGENT Center for Plant Systems Biology Venn diagram tool on http://bioinformatics.psb.ugent.be/webtools/ Venn/ This work was carried out on the Dutch national e-infrastructure with the support of SURF Cooperative. This work was funded by CHDI and Campagneteam Huntington. Funding Information: This work was carried out on the Dutch national e-infrastructure with the support of SURF Cooperative. This work was funded by CHDI and Campagneteam Huntington. Publisher Copyright: © 2021 - The authors. Published by IOS Press.
PY - 2021
Y1 - 2021
N2 - Background: Huntington's disease is a neurodegenerative disorder caused by a CAG expansion in the huntingtin gene, resulting in a polyglutamine expansion in the ubiquitously expressed mutant huntingtin protein. Objective: Here we set out to identify proteins interacting with the full-length wild-type and mutant huntingtin protein in the mice cortex brain region to understand affected biological processes in Huntington's disease pathology. Methods: Full-length huntingtin with 20 and 140 polyQ repeats were formaldehyde-crosslinked and isolated via their N-terminal Flag-tag from 2-month-old mice brain cortex. Interacting proteins were identified and quantified by label-free liquid chromatography-mass spectrometry (LC-MS/MS). Results: We identified 30 interactors specific for wild-type huntingtin, 14 interactors specific for mutant huntingtin and 14 shared interactors that interacted with both wild-type and mutant huntingtin, including known interactors such as F8a1/Hap40. Syt1, Ykt6, and Snap47, involved in vesicle transport and exocytosis, were among the proteins that interacted specifically with wild-type huntingtin. Various other proteins involved in energy metabolism and mitochondria were also found to associate predominantly with wild-type huntingtin, whereas mutant huntingtin interacted with proteins involved in translation including Mapk3, Eif3h and Eef1a2. Conclusion: Here we identified both shared and specific interactors of wild-type and mutant huntingtin, which are involved in different biological processes including exocytosis, vesicle transport, translation and metabolism. These findings contribute to the understanding of the roles that wild-type and mutant huntingtin play in a variety of cellular processes both in healthy conditions and Huntington's disease pathology.
AB - Background: Huntington's disease is a neurodegenerative disorder caused by a CAG expansion in the huntingtin gene, resulting in a polyglutamine expansion in the ubiquitously expressed mutant huntingtin protein. Objective: Here we set out to identify proteins interacting with the full-length wild-type and mutant huntingtin protein in the mice cortex brain region to understand affected biological processes in Huntington's disease pathology. Methods: Full-length huntingtin with 20 and 140 polyQ repeats were formaldehyde-crosslinked and isolated via their N-terminal Flag-tag from 2-month-old mice brain cortex. Interacting proteins were identified and quantified by label-free liquid chromatography-mass spectrometry (LC-MS/MS). Results: We identified 30 interactors specific for wild-type huntingtin, 14 interactors specific for mutant huntingtin and 14 shared interactors that interacted with both wild-type and mutant huntingtin, including known interactors such as F8a1/Hap40. Syt1, Ykt6, and Snap47, involved in vesicle transport and exocytosis, were among the proteins that interacted specifically with wild-type huntingtin. Various other proteins involved in energy metabolism and mitochondria were also found to associate predominantly with wild-type huntingtin, whereas mutant huntingtin interacted with proteins involved in translation including Mapk3, Eif3h and Eef1a2. Conclusion: Here we identified both shared and specific interactors of wild-type and mutant huntingtin, which are involved in different biological processes including exocytosis, vesicle transport, translation and metabolism. These findings contribute to the understanding of the roles that wild-type and mutant huntingtin play in a variety of cellular processes both in healthy conditions and Huntington's disease pathology.
KW - Huntington's disease
KW - cerebral cortex
KW - cross-linking reagents
KW - huntingtin protein
KW - immunoprecipitation
KW - protein interaction mapping
KW - proteomics
UR - http://www.scopus.com/inward/record.url?scp=85112739363&partnerID=8YFLogxK
U2 - https://doi.org/10.3233/JHD-210476
DO - https://doi.org/10.3233/JHD-210476
M3 - Article
C2 - 34151850
SN - 1879-6397
VL - 10
SP - 335
EP - 347
JO - Journal of Huntington's Disease
JF - Journal of Huntington's Disease
IS - 3
ER -