Measuring long‐chain acyl‐coenzyme A concentrations and enrichment using liquid chromatography/tandem mass spectrometry with selected reaction monitoring

Błachnio-Zabielska, Agnieszka; Koutsari, Christina; Jensen, Michael D.

doi:10.1002/rcm.5110

Cited by 41 publications

(34 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…All analytes were monitored as their [M Ϫ H] Ϫ ions. LCFa-CoA esters were estimated using the LC-MS/MS method (9). After extraction in the presence of internal standard (20 ng of heptadecanoyl-CoA), samples were analyzed by UHPLC-ESI-MS/ MS operating in multiple reaction monitoring mode [Waters Acquity UHPLC, C8 UPLC BEH column 2.1 ϫ 150 mm, 1.7 m (Waters, Milford, MA) and TSQ Quantum Ultra triple-quad mass spectrometer (Thermo Fisher Scientific, Waltham, MA)].…”

Section: Methodsmentioning

confidence: 99%

Impact of insulin deprivation and treatment on sphingolipid distribution in different muscle subcellular compartments of streptozotocin-diabetic C57Bl/6 mice

Zabielski

Błachnio-Zabielska

Lanza

et al. 2014

American Journal of Physiology-Endocrinology and Metabolism

Self Cite

View full text Add to dashboard Cite

Insulin deprivation in type 1 diabetes (T1D) individuals increases lipolysis and plasma free fatty acids (FFA) concentration, which can stimulate synthesis of intramyocellular bioactive lipids such as ceramides (Cer) and longchain fatty acid-CoAs (LCFa-CoAs). Ceramide was shown to decrease muscle insulin sensitivity, and at mitochondrial levels it stimulates reactive oxygen species production. Here, we show that insulin deprivation in streptozotocin diabetic C57BL/6 mice increases quadriceps muscle Cer content, which was correlated with a concomitant decrease in the body fat and increased plasma FFA, glycosylated hemoglobin level (%Hb A 1c), and muscular LCFa-CoA content. The alternations were accompanied by an increase in protein expression in LCFa-CoA and Cer synthesis (FATP1/ACSVL5, CerS1, CerS5), a decrease in the expression of genes implicated in muscle insulin sensitivity (GLUT4, GYS1), and inhibition of insulin signaling cascade by Akt␣ and GYS3␤ phosphorylation under acute insulin stimulation. Both the content and composition of sarcoplasmic fraction sphingolipids were most affected by insulin deprivation, whereas mitochondrial fraction sphingolipids remained stable. The observed effects of insulin deprivation were reversed, except for content and composition of LCFa-CoA, CerS protein expression, GYS1 gene expression, and phosphorylation status of Akt and GYS3␤ when exogenous insulin was provided by subcutaneous insulin implants. Principal component analysis and Pearson's correlation analysis revealed close relationships between the features of the diabetic phenotype, the content of LCFa-CoAs and Cers containing C18-fatty acids in sarcoplasm, but not in mitochondria. Insulin replacement did not completely rescue the phenotype, especially regarding the content of LCFa-CoA, or proteins implicated in Cer synthesis and muscle insulin sensitivity. These persistent changes might contribute to muscle insulin resistance observed in T1D individuals. type 1 diabetes; skeletal muscle; mitochondria; ceramide; long-chain fatty acid-coenzyme A METICULOUS GLYCEMIC CONTROL in type 1 diabetic (T1D) individuals can be achieved by administration of prandial, shortacting, and long-acting insulin to mimic ␤-cell secretion in response to varying nutrient levels. Metabolic control in these individuals and stringent adherence to guidelines normalize the plasma glucose and glycosylated hemoglobin levels (%Hb A 1c ) and allow for relatively normal life in T1D. Yet T1D individuals are at greater risk of developing cardiovascular disorders and also known to develop insulin resistance (12, 17). The molecular mechanisms responsible for insulin resistance in T1D subjects are unclear, but variable glucose concentrations (28, 70), advanced glycation end products (62, 73), or desensitization of target tissues by insulin (61) have been proposed. Subcutaneous insulin delivery using an insulin pump partially mimics ␤-cell insulin secretion. However, unlike in nondiabetic individuals with twofold higher hepatic insulin concentration than in ...

show abstract

Section: Methodsmentioning

confidence: 99%

Impact of insulin deprivation and treatment on sphingolipid distribution in different muscle subcellular compartments of streptozotocin-diabetic C57Bl/6 mice

Zabielski

Błachnio-Zabielska

Lanza

et al. 2014

American Journal of Physiology-Endocrinology and Metabolism

Self Cite

View full text Add to dashboard Cite

show abstract

“…The highest amount of propionyl-CoA was also found in the kidney. Saturated acyl-CoAs with a carbon chain length >C 10 were in the highest abundance in the heart. The brain Acyl-CoAs possess structural diversity to include saturated, unsaturated, hydroxylated, and dicarboxyl functional groups in the acyl chain.…”

Section: Methods Validationmentioning

confidence: 99%

“…Interestingly, the kidney has relatively higher amounts of other short-and medium-chain saturated acyl-CoAs (C 3 to C 10 ) compared with other organs tested. Odd carbon number acyl-CoAs, such as heptanoyl-CoA and pentanoyl-CoA, are uncommon acyl-CoAs.…”

Section: Methods Validationmentioning

confidence: 99%

See 1 more Smart Citation

Novel approach in LC-MS/MS using MRM to generate a full profile of acyl-CoAs: discovery of acyl-dephospho-CoAs

Zhang

Berthiaume

et al. 2014

Journal of Lipid Research

View full text Add to dashboard Cite

This article is available online at http://www.jlr.org Acyl-CoAs are a class of important molecules that play essential roles in many physiological processes ( 1 ), such as fatty acid oxidation, lipid synthesis/remodeling, ketone body synthesis, xenobiotic metabolism, and signaling pathways. Classically, acyl-CoAs such as free CoA, acetyl-CoA, and malonyl-CoA are recognized as regulators of metabolic fl ux. The ratio of acetyl-CoA versus free CoA tightly regulates glycolysis and fatty acid oxidation. Malonyl-CoA attenuates fatty acid oxidation by inhibiting acyl-CoA transport into the mitochondrion for oxidation and is utilized in fatty acid synthesis when its concentration is elevated ( 2 ). Many proteins and genes are dynamically regulated by deacylation and acylation via various acyl-CoAs, such as acetyl-CoA, succinyl-CoA, palmitoyl-CoA, etc. ( 3 ). However, acyl-CoAs present in the cell are diverse and may involve molecules beyond fatty acids and their oxidized derivatives. The metabolism of xenobiotics can also lead to the formation of acyl-CoAs, as demonstrated in our previous work on the metabolism of 4-hydroxy acids from C 4 to C 11 in perfused rat liver. The identifi cation of these novel acyl-CoAs extends our understanding of the new catabolic pathways involved in the disposal of 4-hydroxy acids including drugs of abuse and lipid peroxidation products ( 4-8 ).Acyl-CoAs are exclusive to the intracellular metabolites, and the profi le of these biomolecules is indicative of the local metabolic status. Each organ has its specifi c physiological roles with different energetic demands, and therefore, would be expected to exhibit a unique acyl-CoA profi le. The heart preferentially utilizes fatty acids to meet the ATP demand of mechanical contraction. Glucose and/or ketone bodies serve as the primary substrate of the brain for energy

show abstract

“…The analytical column was a reverse-phase Zorbax SB-C8 day-to-day variation, and can detect concentration and enrichment in as little as 20 mg of muscle. Measuring both concentration and isotopic enrichment of DAG and other lipid groups ( 11,12 ) can help to elucidate the contribution of plasma FFA to intramyocellular fatty acid-containing compounds. We believe that this method can be used to elucidate the dynamics of the intramyocellular DAG pool and its relationship to plasma FFA in physiological and pathological states.…”

Section: Liquid Chromatography Conditionmentioning

confidence: 99%

Intramyocellular diacylglycerol concentrations and [U-13C]palmitate isotopic enrichment measured by LC/MS/MS

Błachnio-Zabielska

Jensen

2013

Journal of Lipid Research

Self Cite

View full text Add to dashboard Cite

Recently, more attention has been focused on DAG because of their purported role to induce muscle and liver insulin resistance when present in excess ( 1-4 ). Because of the biological functions of DAG, especially DAG species containing saturated fatty acids, we endeavored to develop a method to precisely and rapidly measure muscle DAG.Most methods for DAG measurement quantify total DAG content. One of the fi rst and common quantitative analysis of DAG was performed by enzymatic analysis using diacylglycerol kinase assay ( 5 ) using labeled ATP 32 . A nonradioactive technique for total DAG quantifi cation uses HPLC analysis following derivatization ( 6 ). The composition of DAG fatty acids can be determined by hydrolysis of DAG and analysis of the liberated and derivatized fatty acid by the means of gas-liquid chromatography with fl ame ionization detector (GLC-FID). Typically, DAG are separated from an extract of total lipids using thin-layer chromatography (TLC) or HPLC. TLC separation requires long processing times, leads to loss of analyte, and is susceptible to sample contamination. Development of mass spectrometry methods has facilitated analysis of DAG by reducing the analysis time, the amounts of consumables, and the likelihood of sample contamination. Diacylglycerols and triacylglycerols are classes of neutral lipids without any permanent charge; therefore, they are usually analyzed by ESI/MS as a sodium [M + Na] + , lithium [M + Li] + , or ammonium adduct that is created spontaneously by adding the appropriate salt to the solvent system ( 7, 8 ) or after the derivatization procedure ( 9, 10 ).Because the potential contribution of circulating FFA to DAG synthesis is unknown, we developed a new method for measuring both concentration and isotopic enrichment of DAG in tissue from animals infused intravenously with uniformly labeled [U-13 C]palmitate. Our method displays excellent inter-and intraassay reproducibility, low

show abstract

Measuring long‐chain acyl‐coenzyme A concentrations and enrichment using liquid chromatography/tandem mass spectrometry with selected reaction monitoring

Cited by 41 publications

References 36 publications

Impact of insulin deprivation and treatment on sphingolipid distribution in different muscle subcellular compartments of streptozotocin-diabetic C57Bl/6 mice

Impact of insulin deprivation and treatment on sphingolipid distribution in different muscle subcellular compartments of streptozotocin-diabetic C57Bl/6 mice

Novel approach in LC-MS/MS using MRM to generate a full profile of acyl-CoAs: discovery of acyl-dephospho-CoAs

Intramyocellular diacylglycerol concentrations and [U-13C]palmitate isotopic enrichment measured by LC/MS/MS

Contact Info

Product

Resources

About