An improved procedure for the purification of plasma membranes from Dictyostelium discoideum

Can J Biochem. 1977 Dec;55(12):1233-6. doi: 10.1139/o77-184.

Abstract

A novel procedure was recently described for the purification of plasma membranes of Dictyostelium discoideum (Gilkes, N. R. & Weeks, G. (1977) Biochim. Biophys. Acta 464, 142-156). Considerable enrichment of plasma membrane marker enzymes was achieved, but since purified mitochondrial and endoplasmic reticulum fractions were unavailable, it was not possible to accurately assess the contamination level of these organelles. We have therefore slightly modified the plasma membrane preparation procedure, improving purification, and have prepared partially purified mitochondrial and endoplasmic reticulum fractions. The data suggest that the contamination of the plasma membranes by endoplasmic reticulum membranes is no greater than 10%, and probably considerably less. No mitochondrial contamination is detectable.

MeSH terms

  • Alkaline Phosphatase / metabolism
  • Cell Fractionation / methods
  • Cell Membrane* / enzymology
  • Centrifugation
  • Centrifugation, Density Gradient
  • Dictyostelium / ultrastructure*
  • Glucose-6-Phosphatase / metabolism
  • Myxomycetes / ultrastructure*
  • NADPH-Ferrihemoprotein Reductase / metabolism
  • Nucleotidases / metabolism
  • Subcellular Fractions / enzymology
  • Succinate Dehydrogenase / metabolism

Substances

  • Succinate Dehydrogenase
  • NADPH-Ferrihemoprotein Reductase
  • Nucleotidases
  • Alkaline Phosphatase
  • Glucose-6-Phosphatase