Biochemical characterization of matrix vesicles from bone and cartilage.
Extracellular matrix vesicles from bone and epiphyseal cartilage of femur and tibia of rats were isolated by collagenase digestion (crude vesicles) and further purified by sucrose gradient centrifugation. Fractions containing cells and membranes were also isolated from the two tissues. The alkaline and acid phosphatase and ATPase activities, as well as protein content of all fractions including crude and purified matrix vesicles, were assayed. The crude vesicles from both tissues demonstrated a high alkaline phosphatase specific activity (5-20 times higher than in the cell fraction). The total enzyme activities and protein content were significantly higher in all fractions from cartilage than those from bone. A major peak of alkaline phosphatase activity and protein content was obtained following the sucrose gradient centrifugation. The position of this peak was similar for both tissues. The specific activity of alkaline phosphatase of purified matrix vesicles was significantly higher in bone than in cartilage. The phosphatase activities from cartilage and bone showed a similar pH dependence and a similar response to metal ions. Of the metal ions tested (Na+, Mg2+, Zn2+, and Ca2+) only Zn2+ (at 5 mM concentration) inhibited significantly the alkaline phosphatase activity of purified matrix vesicles. The electrophoretic profile of purified matrix vesicles showed eight major protein bands common for both tissues. In addition, cartilage vesicles appeared to possess two peptides not found in bone.
D000135:Acid Phosphatase / Q000032:analysis; D000251:Adenosine Triphosphatases / Q000032:analysis; D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D001857:Bone Matrix / Q000032:analysis* / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D006132:Growth Plate / Q000032:analysis* / Q000201:enzymology; D011506:Proteins / Q000032:analysis; D051381:Rats
D011506:Proteins; D000469:Alkaline Phosphatase; D000135:Acid Phosphatase; D000251:Adenosine Triphosphatases