Fine structural localization of nucleoside phosphatase activity in the urinary bladder of the toad

Bartoszewicz, W; Barrnett, Russell J.

doi:10.1016/s0022-5320(64)80033-2

Cited by 40 publications

(14 citation statements)

References 27 publications

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“…The sections were examined in a Phillips EM 300 electron microscope. Magnification 39,900x ies identified ATP-hydrolyzing activities on the luminal membrane as well [1,21]. This limited and in some respects contradictory information on the histochemistry of the toad bladder complicates the task of membrane isolation in this epithelium, based on enzymatic markers during subcellular fractionation.…”

Section: Discussionmentioning

confidence: 99%

Isolation of radio-iodinated apical and basal-lateral plasma membranes of toad bladder epithelium

Rodriguez

Edelman

1979

J. Membrain Biol.

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The apical and basal-lateral plasma membranes of toad bladder epithelium were radio-iodinated with the glucose-glucose oxidase-lactoperoxidase system. The covalently bound radio iodine was used as a marker during subcellular fractionation and membrane isolation. Homogenization conditions that ensured rupture of more than 80% of the cells without substantial nuclear damage were defined by Normarski optics. The nuclei were separated by differential centrifugation and the apical and basal-lateral components were resolved by differential and sucrose density gradient centrifugation. The apical components yielded two radioactive bands that were identified as glycocalyx and plasma membrane labeled with 125I. The basal-lateral components yielded a hetero-disperse pattern made up of at least 3 radioactive bands, but the bulk of the activity of ouabain-sensitive ATPase comigrated with only one of these bands. The mitochondia, identified by assays for cytochrome oxidase and NADH cytochrome c reductase activities, were separated from the radio-iodine labeled by centrifugation in sucrose density gradients under isokinetic conditions. The labeled glycocalyx and the slowly migrating components of basal-lateral labeling were separated from the radio-iodinated membranes by centrifugation at 100,000 x g x 1 hr after removal of the mitochrondria by the isokinetic method. The labeled membranes were then subjected to ultracentrifugation in sucrose density gradients under isopycnic conditions; the basal-lateral membranes containing ouabain-sensitive ATP-ase were well resolved from the apical membranes by this method. These results provide a relatively rapid method of attaining partial purification of the apical and basal-lateral plasma membranes of toad bladder epithelium.

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Section: Discussionmentioning

confidence: 99%

Isolation of radio-iodinated apical and basal-lateral plasma membranes of toad bladder epithelium

Rodriguez

Edelman

1979

J. Membrain Biol.

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“…In the studies of Novikoff et al (13) and Keller (14), the enzyme was localized along the lateral borders of the cell; Bartoszewicz and Barrnett reported luminal localization (15). ADPase activity was also reported by Novikoff et al, and was restricted to the luminal border of the cell.…”

Section: Phosphatase Activitymentioning

confidence: 77%

“…Histochemical studies of the toad bladder (13)(14)(15) have demonstrated a highly active ATPase. In the studies of Novikoff et al (13) and Keller (14), the enzyme was localized along the lateral borders of the cell; Bartoszewicz and Barrnett reported luminal localization (15).…”

Section: Phosphatase Activitymentioning

confidence: 99%

The Isolation of the Membrane of the Toad Bladder Epithelial Cell

Hays

Barland

1966

The Journal of Cell Biology

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There is increasing interest in the structure and composition of cell membranes, and in techniques for their isolation. The following report describes our method for the isolation of the membrane of the toad bladder epithelial cell, the morphology of the isolated membrane, and preliminary determinations of its phosphatase activity. Our interest in isolating the epithelial cell membrane is based on current evidence that the luminal boundary of the cell is the ultimate site of action of vasopressin (I), and that the serosal boundary is associated with active sodium transport (2, 3).

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“…ATPases have been associated with many functional aspects of cell membranes such as ion transport (37), the movement of water (2), and pinocytosis (19). In plants, ATPases have been identified biochemically in mitochondria and chloroplasts, where their activities presumably represent partial reactions of oxidative and photosynthetic phosphorylation.…”

mentioning

confidence: 99%

“…Recent work (18,23,24,35,39) has emphasized the problems inherent in the lead precipitation methods employed in such 'This investigation was supported by a National Science Foundation grant to A. W. Galston. 2 Present address: Botany Department, University of Aberdeen, Aberdeen, Scotland. 3 Present address: Department of Anatomy, Yale University School of Medicine, New Haven, Connecticut 06510.…”

mentioning

confidence: 99%

Localization of Adenosine Triphosphatase Activity on the Chloroplast Envelope in Tendrils of Pisum sativum

1970

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Fine structural localization of nucleoside phosphatase activity in the urinary bladder of the toad

Cited by 40 publications

References 27 publications

Isolation of radio-iodinated apical and basal-lateral plasma membranes of toad bladder epithelium

Isolation of radio-iodinated apical and basal-lateral plasma membranes of toad bladder epithelium

The Isolation of the Membrane of the Toad Bladder Epithelial Cell

Localization of Adenosine Triphosphatase Activity on the Chloroplast Envelope in Tendrils of Pisum sativum

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