Canine traceealis membrane fractionation and characterization

A. K. Grover; M. S. Kannan; E. E. Daniel

doi:10.1016/0143-4160(80)90027-5

Canine traceealis membrane fractionation and characterization

A. K. Grover, M. S. Kannan, E. E. Daniel

Veterinary and Biomedical Sciences

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Abstract

Canine trachealis was homogenized and the various membrane fractions isolated by differential centrifugation and discontinuous sucrose gradient centrifugation. A membrane fraction enriched in the plasma membrane marker enzymes 5′-nucleotidase (5-fold) and K⁺-activated ouabain sensitive p-nitrophenylphosphatase (3-fold) was obtained. The fraction contained very low levels of the inner mitochondrial marker succinate: cytochrome c oxidoreductase. These plasma membrane vesicles showed higher ATP-dependent Ca-uptake (20 μmoles/g protein) than any other submicrosomal fraction. The active Ca-uptake was enhanced by oxalate. The Ca taken up by the plasma membrane vesicles was released instantaneously by dilution in 5mM EGTA and 10μM A23187 and more slowly by dilution only in 5mM EGTA.

Original language	English (US)
Pages (from-to)	135-146
Number of pages	12
Journal	Cell Calcium
Volume	1
Issue number	2
DOIs	https://doi.org/10.1016/0143-4160(80)90027-5
State	Published - Mar 1980

Bibliographical note

Funding Information:
This work was supported by a grant from the Medical Research Council of Canada. The authors wish to thank Dr. T. Jones for help with some experiments.

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10.1016/0143-4160(80)90027-5

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title = "Canine traceealis membrane fractionation and characterization",

abstract = "Canine trachealis was homogenized and the various membrane fractions isolated by differential centrifugation and discontinuous sucrose gradient centrifugation. A membrane fraction enriched in the plasma membrane marker enzymes 5′-nucleotidase (5-fold) and K+-activated ouabain sensitive p-nitrophenylphosphatase (3-fold) was obtained. The fraction contained very low levels of the inner mitochondrial marker succinate: cytochrome c oxidoreductase. These plasma membrane vesicles showed higher ATP-dependent Ca-uptake (20 μmoles/g protein) than any other submicrosomal fraction. The active Ca-uptake was enhanced by oxalate. The Ca taken up by the plasma membrane vesicles was released instantaneously by dilution in 5mM EGTA and 10μM A23187 and more slowly by dilution only in 5mM EGTA.",

author = "Grover, {A. K.} and Kannan, {M. S.} and Daniel, {E. E.}",

note = "Funding Information: This work was supported by a grant from the Medical Research Council of Canada. The authors wish to thank Dr. T. Jones for help with some experiments.",

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T1 - Canine traceealis membrane fractionation and characterization

AU - Grover, A. K.

AU - Kannan, M. S.

AU - Daniel, E. E.

N1 - Funding Information: This work was supported by a grant from the Medical Research Council of Canada. The authors wish to thank Dr. T. Jones for help with some experiments.

PY - 1980/3

Y1 - 1980/3

N2 - Canine trachealis was homogenized and the various membrane fractions isolated by differential centrifugation and discontinuous sucrose gradient centrifugation. A membrane fraction enriched in the plasma membrane marker enzymes 5′-nucleotidase (5-fold) and K+-activated ouabain sensitive p-nitrophenylphosphatase (3-fold) was obtained. The fraction contained very low levels of the inner mitochondrial marker succinate: cytochrome c oxidoreductase. These plasma membrane vesicles showed higher ATP-dependent Ca-uptake (20 μmoles/g protein) than any other submicrosomal fraction. The active Ca-uptake was enhanced by oxalate. The Ca taken up by the plasma membrane vesicles was released instantaneously by dilution in 5mM EGTA and 10μM A23187 and more slowly by dilution only in 5mM EGTA.

AB - Canine trachealis was homogenized and the various membrane fractions isolated by differential centrifugation and discontinuous sucrose gradient centrifugation. A membrane fraction enriched in the plasma membrane marker enzymes 5′-nucleotidase (5-fold) and K+-activated ouabain sensitive p-nitrophenylphosphatase (3-fold) was obtained. The fraction contained very low levels of the inner mitochondrial marker succinate: cytochrome c oxidoreductase. These plasma membrane vesicles showed higher ATP-dependent Ca-uptake (20 μmoles/g protein) than any other submicrosomal fraction. The active Ca-uptake was enhanced by oxalate. The Ca taken up by the plasma membrane vesicles was released instantaneously by dilution in 5mM EGTA and 10μM A23187 and more slowly by dilution only in 5mM EGTA.

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Canine traceealis membrane fractionation and characterization

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