Isolation and properties of skeletal muscle plasma membrane
Biochimica et Biophysica Acta (BBA) - BiomembranesPublished 1 March 1973
A. M. Kidwai, Martha A. Radcliffe, Eda Y. Lee, Edwin E. Daniel
Citations106
SJR quartileQ1
SJR score0.81
SNIP0.77
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TL;DR
This method yielded fairly pure subcellular fractions: namely, plasma membrane, sarcoplasmic reticulum and mitochondria during a short period of time (4 h) without extraction by salts or other drastic procedures.
Abstract
1. Subcellular fractions, namely plasma membrane, sarcoplasmic reticulum and mitochondria were isolated from rat skeletal muscle by the use of a continuous density gradient prepared by mixing 2 M and 0.25 M sucrose in an ISCO density gradient former. 2. The contractile proteins were insoluble under the conditions and were removed by filtration. 3. On examination with the electron microscope the subcellular fractions were fairly homogenous. The unit membrane structure of plasma membrane and sarcoplasmic reticulum were evident at high magnifications, the former being thicker. 4. (Na+ + K+)-ATPase and 5′-nucleotidase (EC 3.1.3.5) were found to be concentration in plasma membrane fraction. Cytochrome c oxidase (EC 1.9.3.1) was measured as a mitochondrial marker and was present in Fraction F3 exclusively. 5. Cholesterol content was highest in plasma membrane fraction and a ratio of cholesterol:phospholipid was found to be 0.11 in plasma membrane, 0.025 in sarcoplasmic reticulum and 0.013 in mitochondria. 6. [3H]Leucine incorporation was maximum in the sarcoplasmic reticulum during a period of 2–5 min. 7. This method yielded fairly pure subcellular fractions: namely, plasma membrane, sarcoplasmic reticulum and mitochondria during a short period of time (4 h) without extraction by salts or other drastic procedures.
Keywords
MedicineBiochemistry, Genetics and Molecular Biology
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