Quantifying acyl-chain diversity in isobaric compound lipids containing monomethyl branched-chain fatty acids.

Autor:	Green CR; Molecular and Cellular Biology Laboratory, The Salk Institute for Biological Studies, 10010N. Torrey Pines Rd., La Jolla, 92037, CA, USA., Kolar MJ; Molecular and Cellular Biology Laboratory, The Salk Institute for Biological Studies, 10010N. Torrey Pines Rd., La Jolla, 92037, CA, USA; Department of Dermatology, University of California, San Diego, La Jolla, CA 92037, USA., McGregor GH; Molecular and Cellular Biology Laboratory, The Salk Institute for Biological Studies, 10010N. Torrey Pines Rd., La Jolla, 92037, CA, USA., Nelson AT; Department of Pathology & Laboratory Medicine, University of Rochester Medical Center, 601 Elmwood Avenue, Rochester, NY 14642., Wallace M; School of Agriculture and Food Science, University College Dublin, Belfield, D04 V1W8, Dublin, Ireland; Conway Institute of Biomolecular and Biomedical Research, Dublin, Ireland., Metallo CM; Molecular and Cellular Biology Laboratory, The Salk Institute for Biological Studies, 10010N. Torrey Pines Rd., La Jolla, 92037, CA, USA. Electronic address: metallo@salk.edu.
Jazyk:	angličtina
Zdroj:	Journal of lipid research [J Lipid Res] 2024 Oct 25, pp. 100677. Date of Electronic Publication: 2024 Oct 25.
DOI:	10.1016/j.jlr.2024.100677
Abstrakt:	Compound lipids comprise a diverse group of metabolites present in living systems, and metabolic- and environmentally-driven structural distinctions across this family is increasingly linked to biological function. However, methods for deconvoluting these often isobaric lipid species are lacking or require specialized instrumentation. Notably, acyl-chain diversity within cells may be influenced by nutritional states, metabolic dysregulation, or genetic alterations. Therefore, a reliable, validated method of quantifying structurally similar even-, odd-, and branched-chain acyl groups within intact compound lipids will be invaluable for gaining molecular insights into their biological functions. Here we demonstrate the chromatographic resolution of isobaric lipids containing distinct combinations of straight-chain and branched-chain acyl groups via ultra-high-pressure liquid chromatography (UHPLC)-mass spectrometry (MS) using a C30 liquid chromatography column. Using metabolically-engineered adipocytes lacking branched-keto acid dehydrogenase A (Bckdha), we validate this approach through a combination of fatty acid supplementation and metabolic tracing using monomethyl branched-chain fatty acids and valine. We observe resolution of numerous isobaric triacylglycerols and other compound lipids, demonstrating the resolving utility of this method. This approach adds to the toolbox for laboratories to quantify and characterize acyl chain diversity across the lipidome. Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article. (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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