Hayashi H, Niinobe S, Matsumoto Y, Suga T
J Biochem. 1981 Feb;89(2):573-9. doi: 10.1093/oxfordjournals.jbchem.a133233.
The characteristics of lipoprotein lipolytic activity in lysosomes after the administration of Triton WR-1339 were studied, and the observed decrease in the density of the particles is discussed. The light mitochondrial fraction prepared from rat liver according to the method of De Duve et al. (Biochem. J. (1955) 60, 604) was used as a crude lysosomal fraction, in which acid phosphatase and lipoprotein lipase activities were concentrated. The lipoprotein lipolytic activity of lysosomes had a pH optimum of 4.0. The activity was strongly inhibited by Triton WR-1339 in vitro at low concentrations, while the acid lipase activity was almost unaffected, though relatively high concentration of the detergent significantly inhibited the latter activity. When Triton WR-1339 administered to rats (150 mg/100 g body weight), the activities of both the lipoprotein lipase and acid lipase of lysosomes from the treated rats decreased to one-third of those of control rats. Low-density and high-density lysosomes were partially purified from the light mitochondrial fraction from Triton WR-1339-treated and silver colloid-treated rats, respectively, by sucrose density gradient centrifugation. The low-density lysosomes (d = 1.00-1.13) from Triton WR-1339 administered rats had approximately 4 and 3 times higher contents of triglyceride and cholesterol, respectively, than the high-density lysosomes (d greater than 1.30) from silver colloid-treated rats. In view of these results and the fact that the density of Triton WR-1339 is quite high (at least d = 1.20), the decrease in density of hepatic lysosomes upon Triton WR-1339 administration cannot be due simply to incorporation of the detergent, and may rather be a result of incorporation and accumulation of some lipid(s) (possibly as lipoprotein) into lysosomes together with Triton WR-1339.
研究了给予Triton WR - 1339后溶酶体中脂蛋白脂解活性的特征,并讨论了观察到的颗粒密度降低的情况。按照De Duve等人(《生物化学杂志》(1955年)60卷,604页)的方法从大鼠肝脏制备的轻线粒体部分用作粗溶酶体部分,其中酸性磷酸酶和脂蛋白脂肪酶活性得以富集。溶酶体的脂蛋白脂解活性的最适pH为4.0。在体外,低浓度的Triton WR - 1339能强烈抑制该活性,而酸性脂肪酶活性几乎不受影响,尽管相对高浓度的去污剂会显著抑制后者的活性。当给大鼠注射Triton WR - 1339(150毫克/100克体重)时,处理组大鼠溶酶体中脂蛋白脂肪酶和酸性脂肪酶的活性均降至对照组大鼠的三分之一。分别通过蔗糖密度梯度离心从Triton WR - 1339处理组和银胶体处理组大鼠的轻线粒体部分中部分纯化出低密度和高密度溶酶体。给予Triton WR - 1339的大鼠的低密度溶酶体(d = 1.00 - 1.13)中甘油三酯和胆固醇的含量分别比银胶体处理组大鼠的高密度溶酶体(d大于1.30)高约4倍和3倍。鉴于这些结果以及Triton WR - 1339的密度相当高(至少d = 1.20)这一事实,给予Triton WR - 1339后肝脏溶酶体密度的降低不能仅仅归因于去污剂的掺入,而可能是某些脂质(可能以脂蛋白形式)与Triton WR - 1339一起掺入并积累到溶酶体中的结果。