人體紅血球葡萄醣六磷酸去氫的純化及性質的研究

Abstract

葡萄醣六磷酸去氫?(Glucose-6-phosphate dehydrogenase, G6PD)是人類紅血球維持正常生理代謝機能時不可缺少的酵素。 我們以N6-(aminohexyl) adenosine 2,5 biphosphate接於Sepharose 4B去做親和柱體(Affinity Chromatography)。可自人體紅血球分離得純質G6PD，以電泳鑑定其純度，亦證明此種分離方法十分良好。 我們於1M urea存在下，用粗製G6PD做等電荷焦點電泳（Isoelectric focusing, IEF），發現至少有七種以上的G6PD band出現。其等電點分別為pH 6.88, 6.79, 6.64, 6.5, 6.39, 6.19, 6.10。再將此七種G6PD band做disc electrophoresis，而發現每一 G6PD band又可分成二個成份，經鑑定而知其中之一分子量為96,000左右，可能是G6PD dimer，另一分子量為50,600-66,800，可能是monomer或是dimer分解物。

A non-equilibrium pH gradient electrophoresis conducted in 6'acrylamide gel in the presence of 1.0 M urea was used to separate isozymes of g1ucose-6-phosphate dehydrogenase in human erythrocytes. At least seven sharp bands were found to vary both in isoelectric point and in the relative intensity. Their isoelectric points, using pH 5-8 range ampholytes, were pH 6.88. for band 1, pH 6.79 for band 2, pH 6.64 for band 3, pH 6.5 for band 4, pH 6.39 for band 5, pH 6.19 for band 6, pH 6.1 for band 7. Each band was resolved further by a disc electrophoresis into two components, the slow(S) and the fast(f) components defined according to their mobilities in the disc gel. All the seven S-component appear to be dimor having molcular weight between 98000-94000. Any one of the seven f-component has a molecular size smaller than the S-component. Their molecular weight were estimated to be between 66800-50600.