Davies P, Drath D B, Engel E E, Huber G L
Lab Invest. 1979 Feb;40(2):221-6.
A combined biochemical and cytochemical study of catalase was performed on alveolar macrophages lavaged from the lungs of adult male rats. Biochemically, catalase activity was present in both a high-speed granule fraction and in the supernatant. The granule-associated activity exhibited latency. Two methods of cell breakage, sonication and homogenization, yielded similar levels and distributions of catalase activity. Catalase activity in whole cells was identified cytochemically by the alkaline diaminobenzidine method and was localized within membrane-lined cytoplasmic granules similar in size to microperoxisomes and associated with cisternae of smooth endoplasmic reticulum. Localization of the reaction product was inhibited by 0.04 M aminotriazole, by cyanide, and by boiling prior to incubation. The cytochemical reaction continued in the absence of exogenous peroxide, but could be prevented by addition of catalase or pyruvate to the peroxide-free medium. Enzyme activity was also localized within a portion of the membrane-bound granules present in the cell fractions used for the biochemical assays.
对成年雄性大鼠肺灌洗得到的肺泡巨噬细胞进行了过氧化氢酶的生物化学和细胞化学联合研究。在生物化学方面，过氧化氢酶活性存在于高速颗粒组分和上清液中。颗粒相关活性表现出潜伏性。两种细胞破碎方法，超声处理和匀浆，产生了相似水平和分布的过氧化氢酶活性。通过碱性二氨基联苯胺法在细胞化学上鉴定了全细胞中的过氧化氢酶活性，其定位于膜内衬的细胞质颗粒内，这些颗粒大小与微过氧化物酶体相似，并与光滑内质网的池相关。反应产物的定位受到0.04 M氨基三唑、氰化物和孵育前煮沸的抑制。在没有外源过氧化物的情况下，细胞化学反应仍在继续，但可以通过向无过氧化物培养基中添加过氧化氢酶或丙酮酸来阻止。酶活性也定位于用于生化分析的细胞组分中存在的一部分膜结合颗粒内。
