Isolation of a plasma membrane-enriched fraction from collagenase-suspended rachitic rat growth plate chondrocytes.
An attempt was made to concentrate plasma membranes of homogenized chondrocytes isolated by collagenase digestion of rachitic rat epiphyseal growth plate cartilage. This study reports the characterization of enzymes in the plasma membrane of isolated chondrocytes and their comparison with extracellular matrix vesicle components. The plasma membrane-enriched fractions that were obtained showed a sevenfold increase in 5'-nucleotidase and a 15-fold increase in alkaline phosphatase, both of which are regarded as plasma membrane markers. SDS-polyacrylamide gel electrophoretic profiles of proteins extracted from membrane fractions contained several major protein bands also seen in isolated matrix vesicles. These studies indicate the usefulness of concentrating plasma membrane components from isolated chondrocytes, after the chondrocytes have been enzymatically freed from investing matrix and other stromal components by collagenase.
D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D002113:Calcification, Physiologic; D002356:Cartilage / Q000032:analysis* / Q000648:ultrastructure; D002462:Cell Membrane / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D005109:Extracellular Matrix / Q000648:ultrastructure; D006132:Growth Plate / Q000032:analysis*; D009708:Nucleotidases / Q000032:analysis; D051381:Rats; D011919:Rats, Inbred Strains; D012279:Rickets / Q000378:metabolism
D009708:Nucleotidases; C018775:adenosine monophosphatase; D000469:Alkaline Phosphatase