Libbin R M, Person P, Gordon A S
Science. 1974 Sep 27;185(4157):1174-6. doi: 10.1126/science.185.4157.1174.
The "light" mitochondrial pellet obtained from the kidneys of rats previously treated with Triton WR-1339 and rendered hypoxic was separated into subcellular component fractions by sucrose density gradient centrifugation in a zonal rotor. Selected fractions were pooled, disrupted by osmotic lysis and repeated freeze-thawing, and incubated in the presence and absence of normal rat serum. The incubation mixtures were assayed for erythropoiesis-stimulating activity (erythropoietin). High specific activity was identified only in fractions rich in lysosomes. Biochemical analysis of reference enzymes for the identification of lysosomes and mitochondria, supplemented by electron microscopic examination of the various separated fractions, supports the observed requirement for lysosomal constituents in the formation of erythropoietin by the kidney.
从预先用曲拉通WR - 1339处理并造成缺氧的大鼠肾脏中获得的“轻”线粒体沉淀，通过在区带转子中进行蔗糖密度梯度离心，分离成亚细胞组分部分。将选定的部分合并，通过渗透裂解和反复冻融使其破碎，并在有和没有正常大鼠血清的情况下进行孵育。对孵育混合物进行促红细胞生成活性（促红细胞生成素）检测。仅在富含溶酶体的部分中鉴定出高比活性。通过对用于鉴定溶酶体和线粒体的参考酶进行生化分析，并辅以对各种分离部分的电子显微镜检查，支持了肾脏在促红细胞生成素形成过程中对溶酶体成分的观察到的需求。
