Emiliani C, Falzetti F, Orlacchio A, Stirling J L
Dipartimento di Medicina Sperimentale e Scienze Biochimiche, Universita di Perugia, Italy.
Biochem J. 1990 Nov 15;272(1):211-5. doi: 10.1042/bj2720211.
beta-N-Acetylhexosaminidase of HL-60 cells was separated into two main forms, A and S, by chromatography on DEAE-cellulose. Analysis of developmental changes in the isoenzyme pattern was complicated by the fact that the specific activity of beta-N-acetylhexosaminidase underwent a 6-fold change during the normal growth cycle. Two other lysosomal enzymes, beta-galactosidase and alpha-mannosidase, behaved similarly. Induction of differentiation of HL-60 cells with dimethyl sulphoxide at a low cell density (3 x 10(5) cells/ml) had a greater effect on the abundance of alpha-subunits of beta-N-acetylhexosaminidase, measured with 4-methylumbelliferyl-beta-N-acetylglucosaminide 6-sulphate, than of beta-subunits, measured with 4-methylumbelliferyl-beta-N-acetylglucosamine, and resulted in an isoenzyme profile in which A and B were the major forms, with the levels of form S greatly decreased.
通过在二乙氨基乙基纤维素上进行色谱分析,HL-60细胞的β-N-乙酰己糖胺酶被分离成两种主要形式,A和S。由于β-N-乙酰己糖胺酶的比活性在正常生长周期中发生了6倍的变化,因此对同工酶模式发育变化的分析变得复杂。另外两种溶酶体酶,β-半乳糖苷酶和α-甘露糖苷酶,表现类似。在低细胞密度(3×10⁵个细胞/毫升)下用二甲基亚砜诱导HL-60细胞分化,对于用4-甲基伞形酮基-β-N-乙酰氨基葡萄糖6-硫酸盐测量的β-N-乙酰己糖胺酶α亚基的丰度,比对用4-甲基伞形酮基-β-N-乙酰氨基葡萄糖测量的β亚基的丰度影响更大,并导致一种同工酶谱,其中A和B是主要形式,S形式的水平大大降低。