γ-谷氨酰转肽酶活性电泳染色新方法

Novel Staining Method for Native-electrophoresis on γ-glutamyltranspeptidase

目的:建立一种新的电泳活性染色方法,以快速地对活性电泳中γ-谷氨酰转肽酶(GGT)进行显色,从而准确鉴定和定位该酶,并可用于该酶的电泳法纯化制备。方法:低温下,样品活性电泳结束后立即将浸有γ-L-谷氨酰-α-萘氨和双甘肽混合底物溶液的滤纸贴在胶面上反应5min,然后再用浸有对氨基苯磺酸和亚硝酸钠混合显色液的滤纸覆盖在基质滤纸上显色。结果:在滤纸上很快显示出一条红色条带,经切胶酶促反应检验确定为GGT,经电泳法和高效液相色谱法确定GGT达到了电泳纯。结论:该法快速、灵敏、简单、直观,为GGT的鉴定和纯化提供了一条新思路。

In the paper, a novel staoning method for native- electrophoresis was established to display γ- glutamyltransptidase （GGT） m glue of naive- PAGE quickly,consequently identify and orient the enzyme exactly, and be applied to purify and produce the enzyme by electrophoresis. Methods： after the GGT samples were native - electrophoresed trader low temperature, a filter paper with mixed substrate of γ- L- glutamyl - α - naphthylamide and glycylglyclne was pasted on the glue immediately to react with GGT for 5 minutes, then another filter paper with solution of P - aminobenzene sulfonic acid and sodium nitrite was affixed on the previous one to display the color. Results： a bright red strip was showed on the paper and confirmed as GGT by the characteristic reaction of GGT, and further the product was validated to up to the electrophoresls purity. Conclusion： this method was fast, sensitive, simple and intuitionistic and provided a new idea to identification and purification of GGT.