A Microspectrophotometric Method for the Determination of Cytochrome Oxidase

Cooperstein, S.J.; Lazarow, Arnold

doi:10.1016/s0021-9258(18)44883-1

Cited by 1,289 publications

(54 citation statements)

References 14 publications

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“…All fractions were frozen for chemical and enzymatic analysis at a later time with the exception of cytochrome-oxidase activity which was measured on the day of cell fractionation. The following markers were assayed in all subcellular fractions by the indicated procedures: Na ϩ /K ϩ ATPase ( 22), alkaline phosphodiesterase (23) and free cholesterol (plasma membrane); cytochrome-oxidase (mitochondria) (24) and N-acetyl-␤ -glucosaminidase (lysosomes) (25); DNA (26) and protein (soluble fraction) (27). Total phospholipid mass (total membrane) was determined in each fraction using a phospholipid phosphorus assay (28).…”

Section: Isolation Of Plasma Membranes From Aortic Smooth Muscle Cellsmentioning

confidence: 99%

Physical effects of cholesterol on arterial smooth muscle membranes: evidence of immiscible cholesterol domains and alterations in bilayer width during atherogenesis

Tulenko

Chen

Mason

et al. 1998

Journal of Lipid Research

167

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Small angle X-ray diffraction was used to examine arterial smooth muscle cell (SMC) plasma membranes isolated from control and cholesterol-fed (2%) atherosclerotic rabbits. A microsomal membrane enriched with plasma membrane obtained from animals fed cholesterol for up to 13 weeks showed a progressive elevation in the membrane unesterified (free) cholesterol:phospholipid (C/PL) mole ratio. Beyond 9 weeks of cholesterol feeding, X-ray diffraction patterns demonstrated a lateral immiscible cholesterol domain at 37 Њ C with a unit cell periodicity of 34 Å coexisting within the liquid crystalline lipid bilayer. On warming, the immiscible cholesterol domain disappeared, and on cooling it reappeared, indicating that the immiscible cholesterol domain was fully reversible. These effects were reproduced in a model C/PL binary lipid system. In rabbits fed cholesterol for less than 9 weeks, lesser increases in membrane C/PL mole ratio were observed. X-ray diffraction analysis demonstrated an increase in membrane bilayer width that correlated with the C/PL mole ratio. This effect was also reproduced in a C/PL binar y lipid system. Taken together, these findings demonstrate that in vivo, feeding of cholesterol causes cholesterol-phospholipid interactions in the membrane bilayer that alter bilayer structure and organization. This interaction results in an increase in bilayer width peaking at a saturating membrane cholesterol concentration, beyond which lateral phase separation occurs resulting in the formation of separate cholesterol bilayer domains. These alterations in structure and organization in SMC plasma membranes may have significance in phenotypic modulation or aortic SMC during early atherogenesis.-Tulenko, T. N., M. Chen, P. E. Mason, and R. P. Mason. Physical effects of cholesterol on arterial smooth muscle membranes: evidence of immiscible cholesterol domains and alterations in bilayer width during atherogenesis.

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Section: Isolation Of Plasma Membranes From Aortic Smooth Muscle Cellsmentioning

confidence: 99%

Physical effects of cholesterol on arterial smooth muscle membranes: evidence of immiscible cholesterol domains and alterations in bilayer width during atherogenesis

Tulenko

Chen

Mason

et al. 1998

Journal of Lipid Research

167

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“…Protein levels and enzyme activities of the individual complexes of the respiratory chain (complex I to IV), succinate: Cytochrome C oxidoreductase (SCC), and citrate synthase (CS; a marker of the number of mitochondria 37 ) in the 600 g supernatant were measured spectrophotometrically, after three freeze–thaw cycles on a KoneLab 20XT analyzer (Thermo Scientific) following standard procedures 38 . These enzyme assays are based on previously described methods 37 , 39 – 42 . All measurements were normalized to both CS activity and protein levels.…”

Section: Methodsmentioning

confidence: 99%

Early-adolescent antibiotic exposure results in mitochondrial and behavioral deficits in adult male mice

Tengeler

Emmerzaal

Geenen

et al. 2021

Sci Rep

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Exposure to antibiotic treatment has been associated with increased vulnerability to various psychiatric disorders. However, a research gap exists in understanding how adolescent antibiotic therapy affects behavior and cognition. Many antibiotics that target bacterial translation may also affect mitochondrial translation resulting in impaired mitochondrial function. The brain is one of the most metabolically active organs, and hence is the most vulnerable to impaired mitochondrial function. We hypothesized that exposure to antibiotics during early adolescence would directly affect brain mitochondrial function, and result in altered behavior and cognition. We administered amoxicillin, chloramphenicol, or gentamicin in the drinking water to young adolescent male wild-type mice. Next, we assayed mitochondrial oxidative phosphorylation complex activities in the cerebral cortex, performed behavioral screening and targeted mass spectrometry-based acylcarnitine profiling in the cerebral cortex. We found that mice exposed to chloramphenicol showed increased repetitive and compulsive-like behavior in the marble burying test, an accurate and sensitive assay of anxiety, concomitant with decreased mitochondrial complex IV activity. Our results suggest that only adolescent chloramphenicol exposure leads to impaired brain mitochondrial complex IV function, and could therefore be a candidate driver event for increased anxiety-like and repetitive, compulsive-like behaviors.

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“…Peroxisomes from 4-5 ml of lambda fraction (80-100 mg protein) were prepared by isopycnic equilibrium centrifugation on 30 ml of continuous gradient of 0 to 50% Nycodenz in homogenizing buffer with 4 ml of 55% Nycodenz in homogenizing buffer as a cushion in a 39-ml tube (29). The gradient fractions were collected from the bottom of the tube and subcellular fractions were identified by their respective marker enzymes: catalase for peroxisomes (30), cytochrome c oxidase for mitochondria (31), and NADPH cytochrome c reductase for microsomes (32).…”

Section: Methodsmentioning

confidence: 99%

Localization of nervonic acid β-oxidation in human and rodent peroxisomes: impaired oxidation in Zellweger syndrome and X-linked adrenoleukodystrophy

Sandhir

Khan

Chahal

et al. 1998

Journal of Lipid Research

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Studies with purified subcellular organelles from rat liver indicate that nervonic acid (C 24:1 ) is ␤ -oxidized preferentially in peroxisomes. Lack of effect by etomoxir, inhibitor of mitochondrial ␤ -oxidation, on ␤ -oxidation of lignoceric acid (C 24:0 ), a peroxisomal function, and that of nervonic acid (24:1) compared to the inhibition of palmitic acid (16:0) oxidation, a mitochondrial function, supports the conclusion that nervonic acid is oxidized in peroxisomes. Moreover, the oxidation of nervonic and lignoceric acids was deficient in fibroblasts from patients with defects in peroxisomal ␤ -oxidation [Zellweger syndrome (ZS) and X-linked adrenoleukodystrophy (X-ALD)]. Similar to lignoceric acid, the activation and ␤ -oxidation of nervonic acid was deficient in peroxisomes isolated from X-ALD fibroblasts. Transfection of X-ALD fibroblasts with human cDNA encoding for ALDP (X-ALD gene product) restored the oxidation of both nervonic and lignoceric acids, demonstrating that the same molecular defect may be responsible for the abnormality in the oxidation of nervonic as well as lignoceric acid. Moreover, immunoprecipitation of activities for acyl-CoA ligase for both lignoceric acid and nervonic acid indicate that saturated and monoenoic very long chain (VLC) fatty acids may be activated by the same enzyme. These results clearly demonstrate that similar to saturated VLC fatty acids (e.g., lignoceric acid), VLC monounsaturated fatty acids (e.g., nervonic acid) are oxidized preferentially in peroxisomes and that this activity is impaired in X-ALD. In view of the fact that the oxidation of unsaturated VLC fatty acids is defective in X-ALD patients, the efficacy of dietary monoene therapy, "Lorenzo's oil," in X-ALD needs to be evaluated.-Sandhir, R., M. Khan, A. Chahal, and I. Singh. Localization of nervonic acid ␤ -oxidation in human and rodent peroxisomes: impaired oxidation in Zellweger syndrome and X-linked adrenoleukodystrophy.

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A Microspectrophotometric Method for the Determination of Cytochrome Oxidase

Cited by 1,289 publications

References 14 publications

Physical effects of cholesterol on arterial smooth muscle membranes: evidence of immiscible cholesterol domains and alterations in bilayer width during atherogenesis

Physical effects of cholesterol on arterial smooth muscle membranes: evidence of immiscible cholesterol domains and alterations in bilayer width during atherogenesis

Early-adolescent antibiotic exposure results in mitochondrial and behavioral deficits in adult male mice

Localization of nervonic acid β-oxidation in human and rodent peroxisomes: impaired oxidation in Zellweger syndrome and X-linked adrenoleukodystrophy

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