[36] Purification of a plasma membrane-bound adenosine triphosphatase from plant roots

Hodges, Thomas K.; Leonard, Robert T.

doi:10.1016/0076-6879(74)32039-3

Cited by 609 publications

(381 citation statements)

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“…ATPase at pH 6.5 (9,10,17), and glucan synthetase at high UDP-glucose (9,37), as well as a high sterol to phospholipid molar ratio (10). The density of plasma membrane vesicles in sucrose gradients was between 1.16 and 1.18 g/cc (9, 10, 15-17, 30, 36, 37) which is similar to the density range reported for plasma membranes from cells of animals (5).…”

supporting

confidence: 65%

“…Isopycnic Centrifugation on Linear Sucrose Gradients. Results with oat roots (9,17) showed that several membrane associated ATPase activities could be distinguished when 13,000g and 13,000 to 80,000g pellets were centrifuged in linear sucrose gradients. To determine whether or not this is the case for corn roots and to determine the location of membrane systems such as Golgi apparatus and endoplasmic reticulum in sucrose gradients, the 13,000g and 13,000 to 80,000g membrane fractions from homogenates of corn roots were centrifuged for 15 hr in linear gradients of 15 to 45% (w/w) sucrose.…”

mentioning

confidence: 99%

“…Previous work on the isolation of plasma membranes from oat roots (9,17) showed that plasma membranes equilibrated on sucrose gradients at a density (1.17 g/cc) near that of mitochondria (1.18 g/cc). Hence, for oat roots, preparation of plasma membranes relatively free of mitochondria required that this organelle be cleared from the homogenate by differential centrifugation prior to sucrose density gradient centrifugation.…”

mentioning

confidence: 99%

“…Procedures for the isolation of the plasmalemma from cells of higher plants have been developed in connection with studies on the site of action of auxin in corn coleoptiles (16), the role of cation-ATPase in ion transport in oat roots (9,10,17), and the involvement of membrane-associated polysaccharide synthetases in cell wall biogenesis in onion stems (36,37). The procedures developed employed differential and sucrose density gradient centrifugation of tissue homogenates to obtain a fraction which was as high as 75% (10) in plasma membranes.…”

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confidence: 99%

“…All manipulations were at 0 to 2 C. The rinsed roots (15-20 g fresh weight) were chopped into small segments and ground in a mortar and pestle without added abrasives as previously described (9,17 For the data in Tables I and II (30). The PACP staining procedure was modified so that incubation in periodate and in phosphotungstic-chromic acid was at 38 C rather than at room temperature.…”

mentioning

confidence: 99%

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Isolation of Plasma Membranes from Corn Roots by Sucrose Density Gradient Centrifugation

Leonard¹,

VanDerWoude²

1976

Plant Physiol.

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An investigation was conducted into the isolation of plasma membrane vesicles from primary roots of corn (Zea mays L., WF9 x M14) by sucrose density gradient centrifugation. Identification of plasma membranes in cell fractions was by specific staining with the periodic-chromic-phosphotungstic acid procedure. Plasma membrane vescles were rich in K+-stimulated ATPase activity at pH 6.5, and equilibrated in linear gradients of sucrose at a peak density of about 1.165 g/cc. It was necessary to remove mitochondria (equilibrium density of 1.18 g/cc) from the homogenate before density gradient centrifugation to minimize mitochondrial contamination of the plasma membrane fraction. Endoplasmic reticulum (NADH-cytochrome c reductase) and Golgi apparatus (latent IDPase) had equilibrium densities in sucrose of about 1.10 g/cc and 1.12 to 1.15 g/cc, respectively. A correlation (r = 0.975) was observed between K+-stimulated ATPase activity at pH 6.5 and the content of plasma membranes in various cell fractions. ATPase activity at pH 9 and cytochrome c oxidase activity were also correlated.A major peak of ATPase activity at pH 6.5 was observed at low density in Ficoll after nonequilibrium centrifugation in a combination Ficollsucrose gradient. Twenty to forty percent of the vesicles in this ATPase fraction stained positively for plasma membranes, and with equilibrium centrifugation the major portion of the ATPase activity shifted to densities in sucrose which were characteristic of plasma membranes. All major vesicular ATPase activities observed in Ficoll or sucrose contained substantial amounts of plasma membranes. For unknown reasons, mitochondria and plasma membranes equilibrated over a broader density range and at lower peak densities in sucrose as a result of equilibrium centrifugation through Ficoll.

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confidence: 65%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Isolation of Plasma Membranes from Corn Roots by Sucrose Density Gradient Centrifugation

Leonard¹,

VanDerWoude²

1976

Plant Physiol.

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A method to obtain right-side-out and sealed plasma membrane vesicles from the filamentous fungus Penicillium simplicissimum

Müller¹,

Zanella²,

Burgstaller³

2001

J. Basic Microbiol.

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Plasma membrane vesicles were purified from the citrate excreting filamentous fungus Penicillium simplicissimum. The hyphae were disrupted by grinding in a glass bead mill and plasma membranes were obtained from a crude membrane fraction by partitioning in an aqueous dextran/polyethylene glycol two‐phase system. The resulting enriched plasma membrane fraction contained only minor contaminations of marker enzymes of intracellular membranes, consisted of more than 90% of right‐side‐out vesicles and was sealed for potassium.

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Peroxisome

et al. 2008

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[36] Purification of a plasma membrane-bound adenosine triphosphatase from plant roots

Cited by 609 publications

References 21 publications

Isolation of Plasma Membranes from Corn Roots by Sucrose Density Gradient Centrifugation

Isolation of Plasma Membranes from Corn Roots by Sucrose Density Gradient Centrifugation

A method to obtain right-side-out and sealed plasma membrane vesicles from the filamentous fungus Penicillium simplicissimum

Peroxisome

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