Emiliani C, Sciarra R, Orlacchio A, Stirling J L
Dipartimento di Medicina Sperimentale e Scienze Biochimiche, Università di Perugia, Italy.
Biochim Biophys Acta. 1990 Mar 1;1037(3):265-73. doi: 10.1016/0167-4838(90)90024-a.
The spleen from a patient with hairy-cell leukaemia had beta-N-acetylhexosaminidase activity that could be resolved into three isoenzymes by chromatography on phenyl boronate agarose. Two of these were the major forms, A and B, found in normal tissues but, in addition, there was an 'extra' form that accounted for 15% of total activity. The 'extra' form hydrolysed the synthetic substrate 4-methylumbelliferyl-beta-N-acetylglucosamine 6-sulphate, indicating the presence of alpha-subunits. It was more acidic than A, was less heat-stable and showed no generation of B on denaturation under a variety of conditions. These findings and the immunoblot (Western blotting) analysis demonstrate that the 'extra' form is entirely composed of alpha-subunits, and most closely resembles S, the residual activity in Sandhoff's disease.
毛细胞白血病患者的脾脏具有β-N-乙酰己糖胺酶活性,通过在苯基硼酸琼脂糖上进行层析可将其分解为三种同工酶。其中两种是正常组织中发现的主要形式,A和B,但此外,还有一种“额外”形式,占总活性的15%。这种“额外”形式可水解合成底物4-甲基伞形酮基-β-N-乙酰氨基葡萄糖6-硫酸盐,表明存在α亚基。它比A更具酸性,热稳定性较差,并且在各种条件下变性时均未显示出B的生成。这些发现以及免疫印迹(蛋白质印迹)分析表明,“额外”形式完全由α亚基组成,并且与桑德霍夫病中的残留活性S最为相似。