Premium
Identifying Proteins of the Murine α7‐Nicotinic Acetylcholine Receptor Interactome
Author(s) -
Mulcahy Matthew,
Hawrot Edward
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.lb507
Subject(s) - interactome , nicotinic acetylcholine receptor , acetylcholine receptor , chemistry , nicotinic agonist , biochemistry , biology , computational biology , receptor , gene
The α7‐Nicotinic acetylcholine receptor (α7‐nAChR) is a ligand‐gated channel widely expressed in vertebrates, and comprises the principal α‐bungarotoxin (α‐bgtx) binding protein in the mammalian central nervous system. The proteins that either interact with the α7‐nAChR or are associated with it in a protein complex can be referred to as the α7‐nAChR interactome. To identify the α7‐nAChR interactome in neural tissue, α‐bgtx‐sensitive protein complexes were isolated from mouse whole brain homogenates using α‐bgtx affinity beads. Proteins were eluted, reduced and alkylated before being digested with trypsin in‐solution and resolved with an Orbitrap Fusion mass spectrometer at Harvard Medical School. We used the ProteoIQ™ software suite for further analysis. Identified proteins from samples isolated using α‐bgtx‐affinity beads and those isolated on beads lacking α‐bgtx were compared. Only proteins unique to the α‐bgtx affinity beads isolation were considered to be α7‐nAChR associated proteins. A total of 171 candidate associated proteins were identified (i.e., with 1% FDR, >90% probability of correct assignment, identified in 2 or more replicates, 0% probability of identification in controls). These results are a continuation of our investigation of α7‐nAChR interacting proteins using the latest technologies in mass spectrometry. The interactome identified here includes proteins which may affect α7‐nAChR expression, localization, or modulation. Further study of these proteins may contribute to a better understanding of how they affect α7‐nAChR function and the role they may have in human disease. This research was supported by NIH 1R21AG038774, 1S10RR027027, NSF EPS‐1004057.