Characterization of Ca<sup>2+</sup> Transport in Purified Endoplasmic Reticulum Membrane Vesicles from <i>Lepidium sativum</i> L. Roots

Buckhout, Thomas J.

doi:10.1104/pp.76.4.962

Cited by 67 publications

(33 citation statements)

References 16 publications

(20 reference statements)

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“…Of further note here was the observation that auxin promoted Ca 21 uptake, whereas cytokinin was inhibitory. Endoplasmic reticulum vesicles were isolated by Buckhout (1984), who demonstrated high affinity Ca 21 accumulation that was not dependent on calmodulin. Finally, attention is directed toward the study of Schumaker and Sze (1985), who carefully isolated and identified two membrane fractions from oat roots.…”

Section: Ca 21 and Microsomesmentioning

confidence: 99%

Calcium: A Central Regulator of Plant Growth and Development

2005

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Section: Ca 21 and Microsomesmentioning

confidence: 99%

Calcium: A Central Regulator of Plant Growth and Development

2005

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“…It is therefore of interest to obtain a complete understanding of mechanisms regulating subcellular Ca2" distribution in plant cells. ATP-dependent transport of Ca2' has been demonstrated in microsomal membrane preparations (9,17,24), in mitochondria (10), and in membrane fractions enriched in endoplasmic reticulum (5,6,8,20) or tonoplast (8,20).…”

mentioning

confidence: 99%

Transport Properties of the Tomato Fruit Tonoplast

Joyce

Cramer²,

Reid³

et al. 1988

Plant Physiol.

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“…Recently, ATPases have reported to be associated with ER (4,5), Golgi membranes (6), and plastid envelopes (15), and they have been partially characterized. The tonoplast-enriched fractions separated after a density gradient are usually contaminated with those endomembranes associated with ATPases.…”

mentioning

confidence: 99%

Isolation and Characterization of Tonoplast from Chilling-Sensitive Etiolated Seedlings of Vigna radiata L.

Yoshida¹,

Kawata²,

Uemura³

et al. 1986

Plant Physiol.

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Tonoplasts were isolated in a high purity from etiolated young seedlings of Vigna radiata L. (mung bean) utilizing a sucrose density gradient system. The excised hypocotyls were homogenized in a sorbitol-buffer system and the 3,600 to 156,000g pellets obtained after the differential centrifugations were suspended in a sorbitol medium and loaded on a linear sucrose density gradient. After centrifugation at 89,000g for 2 hours, tonoplasts were banded at the sample load/sucrose interface. Assessed by electron microscopy and marker enzymes, the purity and the quantity were found to be sufficient for biochemical and biophysical analyses. The tonoplasts were associated with N03-sensitive and vanadate-insensitive ATPase. The tonoplast ATPase was stimulated by proton ionophores such as carbonyl cyanide p-trifluoromethoxyphenylhydrazone and gramicidin D, suggesting a proton-pumping enzyme. In the presence of ATP and Mg2", a proton gradient was formed in the isolated tonoplast vesicles as assessed by fluorescence quenching of quinacrine. The tonoplasts contained several kinds of mannosylated or glycosylated glycoproteins and a major protein (65 kilodaltons) which was unique to the membranes.A great deal of information has been accumulated on the proton ATPase located on the tonoplast from several plants (1,7,19,27). The majority of the publications involve measurements carried out on intact vacuoles isolated by the osmotic lysis of enzymically isolated protoplasts under conditions that preserve the integrity of the tonoplasts (1 1, 26).To get more insight into the biochemical and molecular properties of the tonoplast ATPase and the membrane per se, we need membranes both of high purity and in a large quantity sufficient for analytical uses. For this purpose, tonoplast-enriched fractions are separated from crude microsomal membranes by centrifugation on a linear or discontinuous sucrose density gradient or a dextran density gradient (2,7,8,10,16,18,20).ATPases are known to be associated with wide variety of membrane systems in plant cells. Recently, ATPases have reported to be associated with ER (4, 5), Golgi membranes (6), and plastid envelopes (15), and they have been partially characterized. The tonoplast-enriched fractions separated after a density gradient are usually contaminated with those endomembranes associated with ATPases. Tonoplasts could also be separated in a high purity from the intact vacuoles isolated from enzymically prepared protoplasts following an osmotic lysis. However, there ' Partly supported by Science.is difficulty in getting a large quantity of the tonoplasts.This paper reports the isolation of tonoplast from etiolated seedlings of Vigna radiata by a method in which sorbitol was used as an osmotic stabilizer throughout crude membrane preparation prior to a sucrose density gradient centrifugation. This method has permitted us to isolate tonoplast in a high purity and in a large quantity on subsequent sucrose density gradient.MATERIALS AND METHODS Plant Materials. Seeds of Vigna radiata...

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Characterization of Ca²⁺ Transport in Purified Endoplasmic Reticulum Membrane Vesicles from Lepidium sativum L. Roots

Cited by 67 publications

References 16 publications

Calcium: A Central Regulator of Plant Growth and Development

Calcium: A Central Regulator of Plant Growth and Development

Transport Properties of the Tomato Fruit Tonoplast

Isolation and Characterization of Tonoplast from Chilling-Sensitive Etiolated Seedlings of Vigna radiata L.

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Characterization of Ca2+ Transport in Purified Endoplasmic Reticulum Membrane Vesicles from Lepidium sativum L. Roots

Cited by 67 publications

References 16 publications

Calcium: A Central Regulator of Plant Growth and Development

Calcium: A Central Regulator of Plant Growth and Development

Transport Properties of the Tomato Fruit Tonoplast

Isolation and Characterization of Tonoplast from Chilling-Sensitive Etiolated Seedlings of Vigna radiata L.

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Characterization of Ca²⁺ Transport in Purified Endoplasmic Reticulum Membrane Vesicles from Lepidium sativum L. Roots