Identification of the native Torpedo californica nicotinic acetylcholine receptor's glycan composition after a multi-step sequential purification method using MALDI-ToF MS.

Autor: Maldonado-Hernández R; Department of Biology, Ponce Campus, University of Puerto Rico, Ponce, Puerto Rico, USA.; Molecular Sciences Research Center, University of Puerto Rico, San Juan, Puerto Rico, USA., Quesada O; Molecular Sciences Research Center, University of Puerto Rico, San Juan, Puerto Rico, USA.; Department of Physical Sciences, Río Piedras Campus, University of Puerto Rico, San Juan, Puerto Rico, USA., González-Feliciano JA; Clinical Bioreagent Center, University of Puerto Rico, San Juan, Puerto Rico, USA., Baerga-Ortiz A; Molecular Sciences Research Center, University of Puerto Rico, San Juan, Puerto Rico, USA.; Clinical Bioreagent Center, University of Puerto Rico, San Juan, Puerto Rico, USA.; Department of Biochemistry, Medical Sciences Campus, University of Puerto Rico, San Juan, Puerto Rico, USA., Lasalde-Dominicci JA; Molecular Sciences Research Center, University of Puerto Rico, San Juan, Puerto Rico, USA.; Clinical Bioreagent Center, University of Puerto Rico, San Juan, Puerto Rico, USA.; Department of Biology, Río Piedras Campus, University of Puerto Rico, San Juan, Puerto Rico, USA.; Institute of Neurobiology, University of Puerto Rico Medical Science Campus, San Juan, Puerto Rico, USA.
Jazyk: angličtina
Zdroj: Proteomics [Proteomics] 2024 Jan; Vol. 24 (1-2), pp. e2300151. Date of Electronic Publication: 2023 Oct 30.
DOI: 10.1002/pmic.202300151
Abstrakt: 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.
(© 2023 The Authors. PROTEOMICS published by Wiley-VCH GmbH.)
Databáze: MEDLINE
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