Analysis of neuronal cardiolipin and monolysocardiolipin from biological samples with cyclic ion mobility mass spectrometry

The mitochondrial phospholipid cardiolipin (CL) is essential for proper mitochondrial function and energy production. Cardiolipin has four distinct fatty acid tails with varying expression compositions, resulting in a highly variable tissue-specific distribution of isomer expression. Neuronal cardio...

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Main Authors: Katlynn J. Emaus, Carmen A. Dunbar, Joseph Caruso, Brandon T. Ruotolo, Joseph M. Wider, Thomas H. Sanderson
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-05-01
Series:Frontiers in Physiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fphys.2025.1592008/full
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author Katlynn J. Emaus
Katlynn J. Emaus
Carmen A. Dunbar
Carmen A. Dunbar
Joseph Caruso
Brandon T. Ruotolo
Joseph M. Wider
Joseph M. Wider
Joseph M. Wider
Thomas H. Sanderson
Thomas H. Sanderson
Thomas H. Sanderson
Thomas H. Sanderson
author_facet Katlynn J. Emaus
Katlynn J. Emaus
Carmen A. Dunbar
Carmen A. Dunbar
Joseph Caruso
Brandon T. Ruotolo
Joseph M. Wider
Joseph M. Wider
Joseph M. Wider
Thomas H. Sanderson
Thomas H. Sanderson
Thomas H. Sanderson
Thomas H. Sanderson
author_sort Katlynn J. Emaus
collection DOAJ
description The mitochondrial phospholipid cardiolipin (CL) is essential for proper mitochondrial function and energy production. Cardiolipin has four distinct fatty acid tails with varying expression compositions, resulting in a highly variable tissue-specific distribution of isomer expression. Neuronal cardiolipin has a remarkable variety of subspecies and has recently been used as a biomarker to predict brain injury severity following cardiac arrest and traumatic brain injury. Multiple conditions have been associated with disordered cardiolipin remodeling, including Alzheimer’s disease, Parkinson’s disease, Barth syndrome, and astrocytoma. The clinical relevance of cardiolipin as a biomarker and the importance of the mechanistic role of cardiolipin remodeling in disease emphasize the demand for a reliable and accurate means of the identification and quantification of cardiolipin. In this study, we outline the use of a novel method of cardiolipin analysis using cyclic ion mobility mass spectrometry (cIMS-MS) to isolate and identify cardiolipin subspecies in several biological samples. Furthermore, cIMS-MS established the composition of the cardiolipin profile by individual subspecies across biological samples under basal conditions. Monolysocardiolipin (MLCL), the precursor of mature cardiolipin and a primary diagnostic biomarker of Barth syndrome, was isolated from cardiolipin and identified. The monolysocardiolipin:cardiolipin ratio was quantified in brain samples from tafazzin-knockout (KO) mice, demonstrating accumulation of MLCL and providing direct evidence for the validity of this cIMS-MS methodology through genetic loss-of-function. The novel, multiple-pass feature of cIMS-MS enabled the isolation and amplification of less abundant cardiolipin subspecies in both standards and biological samples. This protocol enables rapid analysis of biological samples, allowing researchers to further dissect the mechanistic role of cardiolipin in injury pathology, with simplified sample preparation and reduced potential for artifact introduction.
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spelling doaj-art-4b3c8b44b2c241bdba797f66374628132025-08-20T02:34:42ZengFrontiers Media S.A.Frontiers in Physiology1664-042X2025-05-011610.3389/fphys.2025.15920081592008Analysis of neuronal cardiolipin and monolysocardiolipin from biological samples with cyclic ion mobility mass spectrometryKatlynn J. Emaus0Katlynn J. Emaus1Carmen A. Dunbar2Carmen A. Dunbar3Joseph Caruso4Brandon T. Ruotolo5Joseph M. Wider6Joseph M. Wider7Joseph M. Wider8Thomas H. Sanderson9Thomas H. Sanderson10Thomas H. Sanderson11Thomas H. Sanderson12Neuroscience Graduate Program, University of Michigan, Ann Arbor, MI, United StatesDepartment of Emergency Medicine, University of Michigan, Ann Arbor, MI, United StatesUniversity of Michigan Biological Mass Spectrometry Core Facility, Ann Arbor, MI, United StatesDepartment of Chemistry, University of Michigan, Ann Arbor, MI, United StatesOlink, Thermo Fisher Scientific, Waltham, MA, United StatesDepartment of Chemistry, University of Michigan, Ann Arbor, MI, United StatesNeuroscience Graduate Program, University of Michigan, Ann Arbor, MI, United StatesDepartment of Emergency Medicine, University of Michigan, Ann Arbor, MI, United StatesThe Max Harry Weil Institute for Critical Care Research and Innovation, University of Michigan, Ann Arbor, MI, United StatesNeuroscience Graduate Program, University of Michigan, Ann Arbor, MI, United StatesDepartment of Emergency Medicine, University of Michigan, Ann Arbor, MI, United StatesThe Max Harry Weil Institute for Critical Care Research and Innovation, University of Michigan, Ann Arbor, MI, United StatesDepartment of Molecular and Integrative Physiology, University of Michigan, Ann Arbor, MI, United StatesThe mitochondrial phospholipid cardiolipin (CL) is essential for proper mitochondrial function and energy production. Cardiolipin has four distinct fatty acid tails with varying expression compositions, resulting in a highly variable tissue-specific distribution of isomer expression. Neuronal cardiolipin has a remarkable variety of subspecies and has recently been used as a biomarker to predict brain injury severity following cardiac arrest and traumatic brain injury. Multiple conditions have been associated with disordered cardiolipin remodeling, including Alzheimer’s disease, Parkinson’s disease, Barth syndrome, and astrocytoma. The clinical relevance of cardiolipin as a biomarker and the importance of the mechanistic role of cardiolipin remodeling in disease emphasize the demand for a reliable and accurate means of the identification and quantification of cardiolipin. In this study, we outline the use of a novel method of cardiolipin analysis using cyclic ion mobility mass spectrometry (cIMS-MS) to isolate and identify cardiolipin subspecies in several biological samples. Furthermore, cIMS-MS established the composition of the cardiolipin profile by individual subspecies across biological samples under basal conditions. Monolysocardiolipin (MLCL), the precursor of mature cardiolipin and a primary diagnostic biomarker of Barth syndrome, was isolated from cardiolipin and identified. The monolysocardiolipin:cardiolipin ratio was quantified in brain samples from tafazzin-knockout (KO) mice, demonstrating accumulation of MLCL and providing direct evidence for the validity of this cIMS-MS methodology through genetic loss-of-function. The novel, multiple-pass feature of cIMS-MS enabled the isolation and amplification of less abundant cardiolipin subspecies in both standards and biological samples. This protocol enables rapid analysis of biological samples, allowing researchers to further dissect the mechanistic role of cardiolipin in injury pathology, with simplified sample preparation and reduced potential for artifact introduction.https://www.frontiersin.org/articles/10.3389/fphys.2025.1592008/fullcardiolipinmonolysocardiolipinmass spectrometrycyclic ion mobility mass spectrometryclinical application
spellingShingle Katlynn J. Emaus
Katlynn J. Emaus
Carmen A. Dunbar
Carmen A. Dunbar
Joseph Caruso
Brandon T. Ruotolo
Joseph M. Wider
Joseph M. Wider
Joseph M. Wider
Thomas H. Sanderson
Thomas H. Sanderson
Thomas H. Sanderson
Thomas H. Sanderson
Analysis of neuronal cardiolipin and monolysocardiolipin from biological samples with cyclic ion mobility mass spectrometry
Frontiers in Physiology
cardiolipin
monolysocardiolipin
mass spectrometry
cyclic ion mobility mass spectrometry
clinical application
title Analysis of neuronal cardiolipin and monolysocardiolipin from biological samples with cyclic ion mobility mass spectrometry
title_full Analysis of neuronal cardiolipin and monolysocardiolipin from biological samples with cyclic ion mobility mass spectrometry
title_fullStr Analysis of neuronal cardiolipin and monolysocardiolipin from biological samples with cyclic ion mobility mass spectrometry
title_full_unstemmed Analysis of neuronal cardiolipin and monolysocardiolipin from biological samples with cyclic ion mobility mass spectrometry
title_short Analysis of neuronal cardiolipin and monolysocardiolipin from biological samples with cyclic ion mobility mass spectrometry
title_sort analysis of neuronal cardiolipin and monolysocardiolipin from biological samples with cyclic ion mobility mass spectrometry
topic cardiolipin
monolysocardiolipin
mass spectrometry
cyclic ion mobility mass spectrometry
clinical application
url https://www.frontiersin.org/articles/10.3389/fphys.2025.1592008/full
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