Improved procedures for the determination of lipid phosphorus by malachite green.

Zhou, Xi; Arthur, Gilbert

doi:10.1016/s0022-2275(20)40776-x

Cited by 146 publications

(26 citation statements)

References 8 publications

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“…Thus, no reliable signal was obtained in biological samples with less than 0.5 nmol of cardiolipin. The cardiolipin concentration, measured by this method, correlated well with conventional phospholipid measurements made by colorimetric determination of phosphate after ashing (27).…”

Section: Discussionsupporting

confidence: 60%

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Microanalysis of cardiolipin in small biopsies including skeletal muscle from patients with mitochondrial disease

Schlame

Shanske

Doty

et al. 1999

Journal of Lipid Research

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Cardiolipin is a specific mitochondrial phospholipid that is present in mammalian tissues in low concentration. To measure cardiolipin in small biopsies from patients with mitochondrial disease, we developed a new technique that can detect subnanomolar levels of well-resolved molecular species, the most abundant of which are tetralinoleoylcardiolipin (L 4 ) and trilinoleoyl-oleoyl-cardiolipin (L 3 O). To this end, a fluorescence-labeled derivative of cardiolipin (2-[naphthyl-1 -acetyl]-cardiolipin dimethyl ester) was formed and analyzed by high performance liquid chromatography. Cardiolipin was measured in skeletal muscle biopsies from 8 patients with mitochondrial disease and in 17 control subjects. In 5 patients with mitochondrial disease, cardiolipin content was higher than normal (2.4-7.0 vs. 0.4-2.2 nmol/ mg protein). In 3 patients with mitochondrial disease, the L 4 /L 3 O ratio was lower than normal (2-4 vs. 4-6). Cardiolipin was also measured in various rat and dog muscle tissues. The L 4 /L 3 O ratio was higher in condensed "muscle" type mitochondria (heart ventricle, skeletal muscle, ratios 4-7) than in orthodox "liver" type mitochondria (liver, smooth muscle, heart auricular appendage, H9c2 myoblasts, ratios 0.4-3), suggesting that the L 4 /L 3 O proportion is important for cristae membrane structure.We concluded that the L 4 /L 3 O ratio is a tissue-specific variable that may change in the presence of mitochondrial disease. The new method is suitable to measure cardiolipin in muscle biopsies in order to estimate concentration of mitochondria. -

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Section: Discussionsupporting

confidence: 60%

“…One enzyme unit was defined as 1 mol citrate formed per minute. Phospholipid concentration was determined by colorometric measurement of phosphate liberated by ashing (27). For statistical presentation, means are given with standard deviation.…”

Section: Other Methodsmentioning

confidence: 99%

Microanalysis of cardiolipin in small biopsies including skeletal muscle from patients with mitochondrial disease

Schlame

Shanske

Doty

et al. 1999

Journal of Lipid Research

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“…Polyphosphate has previously been shown to undergo cleavage by exopolyphosphatases, as well as by promiscuous extracellular phosphatases. , Therefore, to test whether our transporter polyplexes would be useful in vitro and in vivo, we investigated their ability to protect the polyP cargo from enzymatic degradation. Protection from phosphatases was determined using an adaptation of a method reported by Lonrez, where degraded polyP was quantified using a malachite green assay . Briefly, complexes were formed between polyP 22 2a and D 7 :G 7 6 and then incubated with varying concentrations of alkaline phosphatase; then, free PO 4 3– was measured colorimetrically.…”

Section: Resultsmentioning

confidence: 99%

Delivery of Inorganic Polyphosphate into Cells Using Amphipathic Oligocarbonate Transporters

et al. 2018

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Inorganic polyphosphate (polyP) is an often-overlooked biopolymer of phosphate residues present in living cells. PolyP is associated with many essential biological roles. Despite interest in polyP’s function, most studies have been limited to extracellular or isolated protein experiments, as polyanionic polyP does not traverse the nonpolar membrane of cells. To address this problem, we developed a robust, readily employed method for polyP delivery using guanidinium-rich oligocarbonate transporters that electrostatically complex polyPs of multiple lengths, forming discrete nanoparticles that are resistant to phosphatase degradation and that readily enter multiple cell types. Fluorescently labeled polyPs have been monitored over time for subcellular localization and release from the transporter, with control over release rates achieved by modulating the transporter identity and the charge ratio of the electrostatic complexes. This general approach to polyP delivery enables the study of intracellular polyP signaling in a variety of applications.

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“…Lipids were then extracted from each fraction according to the method of Bligh and Dyer (1959) and were separated on a silica gel 60 TLC plate (Merck; Darmstadt, Germany) using chloroform/methanol/acetic acid/water (100:75:7:4, v/v). Lipids were then visualized with iodine vapor and compared with those of standards before being assayed for phosphorous content with malachite green reagent (Sigma) (Zhou and Arthur 1992).…”

Section: Methodsmentioning

confidence: 99%

Sphingomyelin Levels in the Plasma Membrane Correlate with the Activation State of Muscle Satellite Cells

Nagata

Kobayashi

Umeda

et al. 2006

J Histochem Cytochem.

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S U M M A R Y Satellite cells are responsible for postnatal growth, hypertrophy, and regeneration of skeletal muscle. They are normally quiescent, and must be activated to fulfill these functions, yet little is known of how this is regulated. As a first step in determining the role of lipids in this process, we examined the dynamics of sphingomyelin in the plasma membrane. Sphingomyelin contributes to caveolae/lipid rafts, which act to concentrate signaling molecules, and is also a precursor of several bioactive lipids. Proliferating or differentiated C2C12 muscle cells did not bind lysenin, a sphingomyelin-specific binding protein, but noncycling reserve cells did. Quiescent satellite cells also bound lysenin, revealing high levels of sphingomyelin in their plasma membranes. On activation, however, the levels of sphingomyelin drop, so that lysenin did not label proliferating satellite cells. Although most satellite cell progeny differentiate, others stop cycling, maintain Pax7, downregulate MyoD, and escape immediate differentiation. Importantly, many of these Pax7-positive/ MyoD-negative cells also regained lysenin binding on their surface, showing that the levels of sphingomyelin had again increased. Our observations show that quiescent satellite cells are characterized by high levels of sphingomyelin in their plasma membranes and that lysenin provides a novel marker of myogenic quiescence. (J Histochem Cytochem 54:375-384, 2006)

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Improved procedures for the determination of lipid phosphorus by malachite green.

Cited by 146 publications

References 8 publications

Microanalysis of cardiolipin in small biopsies including skeletal muscle from patients with mitochondrial disease

Microanalysis of cardiolipin in small biopsies including skeletal muscle from patients with mitochondrial disease

Delivery of Inorganic Polyphosphate into Cells Using Amphipathic Oligocarbonate Transporters

Sphingomyelin Levels in the Plasma Membrane Correlate with the Activation State of Muscle Satellite Cells

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