Hosokawa R, Uchida Y, Fujiwara S, Noguchi T
Department of Prosthetic Dentistry, Kyushu Dental College, Kitakyushu, Japan.
J Biol Chem. 1988 Jul 25;263(21):10045-7.
Matrix vesicles were isolated from epiphyseal growth plates of young rabbits. Lactate dehydrogenase activity was detected in the isolated matrix vesicles only in the presence of detergents, suggesting that NADH, the cofactor for the assay, does not penetrate the membrane of matrix vesicles. In contrast, the activity of alkaline phosphatase, a marker enzyme of the outer surface of matrix vesicles, was detected in the matrix vesicles using p-nitrophenyl phosphate as the substrate both in the presence and absence of detergents. Lactate dehydrogenase activity was detected only in the cytosol of chondrocytes of the epiphyseal growth plates but not in other subcellular fractions, showing that lactate dehydrogenase is not from the plasma membrane and membranes of intracellular organelles of chondrocytes. The isolated matrix vesicles contained all five lactate dehydrogenase isoenzymes but did not possess other cytosolic enzymes. These results show that lactate dehydrogenase is located in the matrix vesicles and suggest the presence of a mechanism for the specific uptake of cytosolic lactate dehydrogenase and the possibility of enzymatic quantification of the matrix vesicles at various calcification sites.
从幼年兔子的骨骺生长板中分离出基质小泡。仅在存在去污剂的情况下,在分离出的基质小泡中检测到乳酸脱氢酶活性,这表明该检测的辅助因子NADH不能穿透基质小泡的膜。相比之下,使用对硝基苯磷酸作为底物,无论有无去污剂,在基质小泡中均检测到碱性磷酸酶的活性,碱性磷酸酶是基质小泡外表面的标记酶。乳酸脱氢酶活性仅在骨骺生长板软骨细胞的胞质溶胶中检测到,而在其他亚细胞组分中未检测到,这表明乳酸脱氢酶并非来自软骨细胞质膜和细胞内细胞器的膜。分离出的基质小泡含有所有五种乳酸脱氢酶同工酶,但不具备其他胞质酶。这些结果表明乳酸脱氢酶位于基质小泡中,并提示存在一种特异性摄取胞质乳酸脱氢酶的机制,以及在不同钙化位点对基质小泡进行酶定量的可能性。