Identification of the native Torpedo californica nicotinic acetylcholine receptor's glycan composition after a multi‐step sequential purification method using MALDI‐ToF MS
Year: 2023
Type: article
Abstract: The Cys-loop pentameric ligand-gated ion channels comprise a dynamic group of proteins that have been extensively studied for decades, yielding a wealth of findings at both the structural and functional levels. The nicotinic acetylcholine receptor (nAChR) is no exception, as it is part of this large protein family involved in proper organismal function. Our efforts have successfully produced a highly pure nAChR in detergent complex (nAChR-DC), enabling more robust studies to be conducted on it, including beginning to experiment with high-throughput crystallization. Our homogeneous product has been identified and extensively characterized with 100% identity using Nano Lc MS/MS and MALDI ToF/ToF for each nAChR subunit. Additionally, the N-linked glycans in the Torpedo californica-nAChR (Tc-nAChR) subunits have been identified. To study this, the Tc-nAChR subunits were digested with PNGase F and the released glycans were analyzed by MALDI-ToF. The MS results showed the presence of high-mannose N-glycan in all native Tc-nAChR subunits. Specifically, the oligommanose population Man8-9GlcNac2 with peaks at m/z 1742 and 1904 ([M + Na]+ ions) were observed.
Source: PROTEOMICS
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $4,220
Awards: Establishment of a multi-user high-performance MALDI ToF/ToF mass spectrometer in the University of Puerto Rico, Molecular Science Research Center, G4986506763, COBRE Phase 2: Center for Neuroplasticity at the University of Puerto Rico, G6005806052, A lipid-based approach towards a high-resolution structure of a functional nAChR, Support for University Biomedical Excellence at UPR-RP, G9000043731, Advancing Competitive Biomedical Research in Puerto Rico, Center for Collaborative Research in Health Disparities, Biology with X-ray Lasers
OpenAlex ID: W4388034607