Oshima G, Yamada M
School of Pharmaceutical Sciences, Kitasato University, Tokyo, Japan.
Biol Chem Hoppe Seyler. 1991 Oct;372(10):947-53. doi: 10.1515/bchm3.1991.372.2.947.
The subcellular distribution of the elastase in human myeloid leukemia HL-60 cells was studied in comparison with that in normal leukocytes. On differential centrifugation, most of the elastase activity of HL-60 cell lysates was recovered in the 105,000 x g supernatant, while that of human peripheral blood leukocyte lysates was recovered in the 500 x g precipitate (azurophil granule-rich fraction). Moreover, on Percoll density gradient centrifugation, the elastase activity in HL-60 cell extracts was recovered in the lightest fraction with none in the azurophil granule-rich fractions, whereas most of the activity in leukocyte extracts was recovered in the azurophil granule-rich fractions. This subcellular localization of elastase did not change when HL-60 cells differentiated into monocytes and granulocytes by induction with 12-O-tetradecanoyl phorbol-13-acetate and retinoic acid, respectively. Furthermore, on Sephadex G-75 gel filtration, the elastase activity in HL-60 cell extracts was eluted earlier than that in leukocyte extracts. The size estimation indicated that the elastase of HL-60 cells was 36-30 kDa, corresponding to the size of an elastase precursor reported. The relevance of a large form of the elastase in HL-60 cells to its subcellular localization is discussed.
对人髓性白血病HL - 60细胞中弹性蛋白酶的亚细胞分布进行了研究,并与正常白细胞中的情况作比较。差速离心时,HL - 60细胞裂解物的大部分弹性蛋白酶活性存在于105,000×g的上清液中,而人外周血白细胞裂解物的弹性蛋白酶活性则存在于500×g的沉淀物(富含嗜天青颗粒的部分)中。此外,在Percoll密度梯度离心时,HL - 60细胞提取物中的弹性蛋白酶活性存在于最轻的部分,富含嗜天青颗粒的部分没有活性,而白细胞提取物中的大部分活性则存在于富含嗜天青颗粒的部分。当HL - 60细胞分别用12 - O - 十四烷酰佛波醇 - 13 - 乙酸酯和视黄酸诱导分化为单核细胞和粒细胞时,弹性蛋白酶的这种亚细胞定位没有改变。此外,在Sephadex G - 75凝胶过滤中,HL - 60细胞提取物中的弹性蛋白酶活性比白细胞提取物中的洗脱得早。大小估计表明,HL - 60细胞的弹性蛋白酶为36 - 30 kDa,与报道的弹性蛋白酶前体大小相对应。文中讨论了HL - 60细胞中这种大分子形式的弹性蛋白酶与其亚细胞定位的相关性。