Clark S A, Ambrose W W, Anderson T R, Terrell R S, Toverud S U
Department of Pharmacology, University of North Carolina, Chapel Hill 27599.
J Bone Miner Res. 1989 Jun;4(3):399-405. doi: 10.1002/jbmr.5650040315.
The intracellular localization of the tartrate-resistant purple acid phosphatase in osteoclasts of developing rat bone has been determined immunocytochemically using an antiserum to the purified bone-derived purple acid phosphatase. The localization of the immunoreactivity was compared with the results of enzyme histochemistry using p-nitrophenylphosphate as substrate and 10 mM tartrate. Both methods revealed the presence of the enzyme in numerous vesicles of various sizes up to 2-3 microns in diameter and in granules. There was no immunoreactivity in the Golgi apparatus, and tartrate completely inhibited the histochemical activity of this organelle. No consistent extracellular activity could be detected, nor was any reaction product observed at the ruffled border. The localization of the tartrate-resistant purple acid phosphatase in osteoclasts is consistent with an intracellular function for this enzyme.
利用针对纯化的骨源性紫色酸性磷酸酶的抗血清,通过免疫细胞化学方法确定了发育中大鼠骨骼破骨细胞中抗酒石酸紫色酸性磷酸酶的细胞内定位。将免疫反应性的定位结果与以对硝基苯磷酸为底物、10 mM酒石酸的酶组织化学结果进行了比较。两种方法均显示该酶存在于直径达2 - 3微米的各种大小的众多囊泡以及颗粒中。高尔基体中没有免疫反应性,并且酒石酸完全抑制了该细胞器的组织化学活性。未检测到一致的细胞外活性,在皱褶缘也未观察到任何反应产物。破骨细胞中抗酒石酸紫色酸性磷酸酶的定位与该酶的细胞内功能一致。