Respiration-dependent accumulation of inorganic phosphate and Ca++ by rat liver mitochondria

Lehninger, Albert L.; Rossi, C; Greenawalt, John W.

doi:10.1016/0006-291x(63)90377-2

Cited by 186 publications

(66 citation statements)

References 5 publications

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“…Furthermore the findings are consistent with the fact that the deposited salt has the C a : P ratio of hydroxyapatite, as reported earlier (3)(4)(5).…”

Section: Discussionsupporting

confidence: 92%

“…The "active" or respiration-linked accumulation of Ca ++ from the suspending medium by isolated mitochondria, first demonstrated by Vasington and Murphy (1,2), has been studied in some detail in a number of recent investigations (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11). The accumulation of Ca ++ during respiration has been found to be quite massive, attaining values as high as 3.0/~moles Ca++ per mg mitochondrial protein (2)(3)(4)(5)(6).…”

Section: Introductionmentioning

confidence: 99%

“…The accumulation of Ca ++ during respiration has been found to be quite massive, attaining values as high as 3.0/~moles Ca++ per mg mitochondrial protein (2)(3)(4)(5)(6). It is accompanied by uptake of inorganic phosphate from the medium; the molar ratio Ca:P of the accumulated salt is about 1.67, or approximately that of hydroxyapatite (4,5).…”

Section: Introductionmentioning

confidence: 99%

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Effect of Active Accumulation of Calcium and Phosphate Ions on the Structure of Rat Liver Mitochondria

Greenawalt

Rossi

Lehninger

1964

The Journal of Cell Biology

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356

126

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Rat liver mitochondria allowed to accumulate maximal amounts of Ca ++ and HPO4 = ions from the suspending medium in vitro during respiration have a considerably higher specific gravity than normal mitochondria and may be easily separated from the latter by isopycnic centrifugation in density gradients of sucrose or cesium chloride. When the mitochondria are allowed to accumulate less than maximal amounts of Ca++ and HPO4= from the medium, they have intermediate specific gravities which are roughly proportional to their content of calcium phosphate. Maximally "loaded" mitochondria are relatively homogeneous with respect to specific gravity. Correlated biochemical and electron microscopic studies show that Ca++-loaded mitochondria contain numerous dense granules, of which some 85 per cent are over 500 A in diameter. These granules are electron-opaque not only following fixation and staining with heavy metal reagents, but also following fixation with formaldehyde, demonstrating that the characteristic granules in Ca++-loaded mitochondria have intrinsic electron-opacity. The dense granules are almost always located within the inner compartment of the mitochondria and not in the space between the inner and outer membranes. They are frequently located at or near the cristae and they often show electrontransparent "cores." Such granules appear to be made up of clusters of smaller dense particles, but preliminary x-ray diffraction analysis and electron diffraction studies have revealed no evidence of crystallinity in the deposits. The electron-opaque granules decrease in number when the Ca++-loaded mitochondria are incubated with 2,4-dinitrophenol; simultaneously there is discharge of Ca ++ and phosphate from the mitochondria into the medium.

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“…Furthermore the findings are consistent with the fact that the deposited salt has the C a : P ratio of hydroxyapatite, as reported earlier (3)(4)(5).…”

Section: Discussionsupporting

confidence: 92%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Effect of Active Accumulation of Calcium and Phosphate Ions on the Structure of Rat Liver Mitochondria

Greenawalt

Rossi

Lehninger

1964

The Journal of Cell Biology

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356

126

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“…Energy can be supplied by either substrate oxidation or ATP. The antibiotic, oligomycin, inhibits ATP-driven Ca uptake, but not substrate-driven uptake (6,18 …”

mentioning

confidence: 99%

Calcium Accumulation by Maize Mitochondria

Hodges

Hanson

1965

Plant Physiol.

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A number of laboratories have reported that animal mitochondria can actively accumulate various inorganic ions (8). Energy can be supplied by either substrate oxidation or ATP. The antibiotic, oligomycin, inhibits ATP-driven Ca uptake, but not substrate-driven uptake (6,18 Materials and MethodsIsolation of Mitochondria. Corn seeds (Zea mays L., WF9 X M14) were germinated in the dark at 28°on paper towels saturated with 10-4 M CaC2.After 3 and one-half days, the shoots were excised, chilled, and ground in an ice-cold mortar with 0.25 M sucrose, 0.05 M KH2PO4 and 0.005 M EDTA, adjusted to pH 7.5 with Tris (hydroxymethyl) aminomethane. The slurry was strained through cheesecloth. Mitochondria were isolated in a refrigerated centrifuge as the fraction sedimenting between 2000 X g for 5 minutes and 12,000 X g for 10 minutes. The mitochondria were twice washed, first in the grinding medium, next in 0.25 M sucrose and were finally suspended in 0.25 M sucrose. The mitochondria were quite active, giving QO2(N) values of about 1500 and P/O values of about 2.5 when oxidizing a mixture of pyruvate and succinate in the absence of inhibitors or uncouplers. Procedures for determining oxidative phosphorylation have been described (12

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“…The ATP-Pi exchange activity was sensitive to DNF and to oligomicin (Table 4) suggesting that this activity was related to oxidative phosphorylation. Accumulation of 45Ca2+ by N. crassa mitochondria was supported by substrate oxidation, but the rate of uptake was considerably slower than that observed with intact rat liver mitochondria (Lehninger, Rossi & Greenawalt, 1963;. This activity, as in rat liver mitochondria, appeared to be respiration-dependent since KCN inhibited the reaction (Table 4).…”

Section: Electron Microscopymentioning

confidence: 83%

The Preparation and Biochemical Properties of Mitochondria from Neurospora crassa

Hall¹,

Greenawalt²

1967

Journal of General Microbiology

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SUMMARYMitochondria were prepared in large quantities from Neurospora crassa by grinding hyphae with glass beads in an Eppenbach Micro-mill. Observation in the electron microscope indicated that some of the isolated mitochondria were probably damaged during preparation. Nevertheless, the crude mitochondrial fraction was capable of coupling phosphorylation to the oxidation of seven different substrates tested. The P:O ratio obtained with succinate ranged from 0.7 to 1-3 in different experiments with different mitochondria1 preparations, but only low respiratory control ratios were observed. The effects of common inhibitors of oxidative phosphorylation were similar to those reported with mammalian mitochondria. In addition, ATPase and ATP-Pi exchange activities, as well as ion accumulation, were measured in the Neurospora mitochondria. The use of a ' preparation ' medium containing sucrose, 0.25 M; EDTA, 0.005 M (pH 7.0); bovine serum albumin (0.15 %, w/v) was necessary to obtain these results.

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Respiration-dependent accumulation of inorganic phosphate and Ca++ by rat liver mitochondria

Cited by 186 publications

References 5 publications

Effect of Active Accumulation of Calcium and Phosphate Ions on the Structure of Rat Liver Mitochondria

Effect of Active Accumulation of Calcium and Phosphate Ions on the Structure of Rat Liver Mitochondria

Calcium Accumulation by Maize Mitochondria

The Preparation and Biochemical Properties of Mitochondria from Neurospora crassa

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