Isolation and purification of normal and malignant colonic plasma membranes

Plasma membrane fractions from normal colon cells and a transplantable colon adenocarcinoma were isolated and purified by differential and zonal density centrifugation. Enrichment of normal and adenocarcinoma plasma membranes was found in zonal fractions I and II (ZI and ZII) following centrifugation in an 18--50% sucrose gradient. The distribution of various marker enzymes in normal colon preparations suggested an apical origin for the membranes obtained in zonal fraction I while zonal fraction II appeared to contain basal-lateral membrane fragments. Enzymatic analysis of the plasma membrane derived from the colon tumor indicated that these fractions possess a more uniform distribution of Na-K+ ATPase perhaps reflecting a dedifferentiated state. The plasma membrane fractions isolated should prove useful for investigation of transport and other properties of vesicles derived from malignant and normal colon cells.