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Analytical subcellular fractionation of rat pituitary homogenates with special reference to the subcellular localization and properties of alkaline phosphatases.
- Source :
-
Biochimica et biophysica acta [Biochim Biophys Acta] 1983 Jun 09; Vol. 757 (3), pp. 296-301. - Publication Year :
- 1983
-
Abstract
- Alkaline phosphatase activities of the virgin rat anterior pituitary were studied with a highly sensitive fluorometric assay. Tissue whole homogenates were fractionated on sucrose density gradients in a Beaufay automatic zonal rotor and the gradient fractions assayed for alkaline phosphatase, prolactin and various organelle marker enzymes. Alkaline phosphatase was distributed between two peaks on the gradient. The low-density (1.10-1.15 g . cm-3) alkaline phosphatase component co-sedimented with the plasma membrane marker, 5'-nucleotidase, had an apparent Km for 4-methylumbelliferyl phosphate of approx. 59 microM, and was inhibited by levamisole. The high-density (1.20-1.25 g . cm-3) peak was resistant to levamisole-inhibition, had an apparent Km of approx. 30 microM and its distribution was distinct from plasma membrane, Golgi, lysosome, endoplasmic reticulum, mitochondria and prolactin granule markers on the isopycnic gradients.
- Subjects :
- Alkaline Phosphatase antagonists & inhibitors
Alkaline Phosphatase classification
Animals
Cell Fractionation methods
Cell Membrane enzymology
Centrifugation, Density Gradient
Chemical Phenomena
Chemistry
Female
Levamisole pharmacology
Prolactin isolation & purification
Rats
Rats, Inbred Strains
Spectrometry, Fluorescence
Subcellular Fractions enzymology
Alkaline Phosphatase isolation & purification
Pituitary Gland enzymology
Subjects
Details
- Language :
- English
- ISSN :
- 0006-3002
- Volume :
- 757
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- Biochimica et biophysica acta
- Publication Type :
- Academic Journal
- Accession number :
- 6849978
- Full Text :
- https://doi.org/10.1016/0304-4165(83)90054-5