人類紅血球葡萄糖六磷酸去氫同功?的鑑定

Abstract

人類紅血球中的葡萄糖六磷酸去氫?(G6PD)有許多同功?之事已被許多前人研究。於IMUrea存在下的6﹪Acrylarmide膠體中進行等電荷焦點電泳(Isoelectric Focusing IEF)6小時，可得到解析度極佳的7個以上G6PD色帶，其等電點分別為PH6.56，6.38，6.21，6.06，5.92，5.80及5.70此與無Urea之IEF所得PH範圍相同。又經由雙向電泳分析，得知上述7個以上色帶並非均勻相，均含少量Monomer或Partial Digest Dimer，大部仍為雙體(Dimer)，故為電荷同功?(Charge Isoenzyme) 利用0.75 M NaCl萃取出G6PD，此乃微量分離鑑定時很好的方法，以此鑑定不同血源及不同老化程度紅血球，均有上述色帶，但色帶之深度，比例也許各有差異。

An improved technique of isoelectric focusing, which is performed in 6.0% polyacrylamide gel in the presence of 1.0 M urea and conducted at 300 V for 3 h and 400 V for another 3 h, separates well the various molecular forms of glucose-6-phosphate dehydrogenase from human erythrocytes. At least seven sharp bands appear in the electrophoretic pattern, which vary both in isoelectric point and in the relative intensity. Their isoelectric points are at pH 6.56 for band 1, pH 6.38 for band 2, pH 6.21 for band 3, pH 6.06 for band 4, pH 5.92 for band 5, pH 5.80 for hand 6, and pH 5.70 for band 7. These seven bands are formed in healthy male bloods. They are present in both young and aged erythrocytes. A second-dimensional disc electrophoresis performed in a polyacrylamide gel slab resolves each band into two components, a slow and a fast component defined according to their mobilities'in the disc gel. The slow component constitutes the major portion of each band. All the seven slow components appear to be dimer having molecular weight between 98,000-94,000. They belong to “charge isomers” having identical molecular size but containing different net charges. The molecular weight of the fast components is between 66,800-50,600. These fast components might be monomer or the digested products of slow components.