杨金奎 陈家伟 何戎华 210029 南京医科大学第一附属医院内分泌科 中华内分泌代谢杂志 1999 0 15 4


    关键词：谷氨酸脱羧酶；纯化；糖尿病，1型；凝胶层析 期刊 zhnfmdxzz 0 论著 fur -->


    

【摘要】 目的 从猪脑中提取纯化谷氨酸脱羧酶(GAD)，用于1型糖尿病患者GAD抗体的测定。方法 以猪脑制备GAD粗酶，经SephadexG100、DEAE-Sephacel和羟基磷灰石凝胶层析分离纯化，结合十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)测定相对分子质量、同位素法测定GAD酶活性，建立了GAD纯化方法。结果 GAD粗酶经纯化后的单位蛋白量的酶活性增加了118倍。经SDS-PAGE分析，为一条相对分子质量约64 000的蛋白条带。结论 用多步层析分离法从猪脑中提取纯化GAD是一种简便可行的方法。

Purification andidentification of glutamic acid decarboxylase from porcine brain

YANG Jinkui,CHEN Jiawei,HERonghua.

Department of Endocrinology,theFirst Affiliated Hospital,Nanjing Medical University,Nanjing,210029

【 Abstract 】 Objective To purify glutamic acid decarboxylase (GAD) from porcinebrain for the sake of detecting GAD antibodies in patients with type 1 diabetes mellitus. Methods GAD was obtained from porcinebrain as a crude extract and purified by using Sephadex G100, DEAE-Sephacel and calciumphosphate chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and GAD activity assay were adopted to identify this purified enzyme. Results With this method, specificactivity of GAD increased by 118-fold and it consisted of a single band around 64000 onSDS-PAGE. Conclusion It is a simple and useful method to purify GAD fromporcine brain by multi-step chromatography.

     【 Keywords 】 Glutamicacid decarboxylase Purification Diabetes mellitus, type 1 Gel chromatography

1型糖尿病是一种自身免疫性疾病 ......

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