Pérez-de la Mora M, Rizo-Silva A B, Méndez-Franco J
Departamento de Neurociencias, Universidad Nacional Autónoma de México, D.F.
Neurochem Res. 1992 Apr;17(4):339-43. doi: 10.1007/BF00974575.
Gel-filtration chromatography from crude extracts of mouse brain indicates the presence of a high molecular weight (HMW) (more than 750 kd) and a low molecular weight (LMW) (87.5 Kd) glutamic acid decarboxylase (GAD) when they are concentrated prior chromatography either by precipitation with salts or ethanol. Kinetically both peaks of GAD-activity exhibit an almost identical Km for L-glutamate. Only LMW-GAD appears if the concentration step is carried out by ultrafiltration or if the extract is chromatographied without the concentrating step. HMW-GAD disappears from the chromatographic profiles if ethanolic extracts of GAD are treated with Triton X-100 before the chromatographic procedure. The sodium sulfate precipitation of a previously separated LMW-GAD gives rise to the reappearance of a HMW-GAD peak. Apparently HMW-GAD does not exist as a different molecular entity; indeed it may be an artefactual aggregation of LMW-GAD.
从小鼠脑粗提物进行的凝胶过滤层析表明,当通过盐析或乙醇沉淀在层析前进行浓缩时,存在高分子量(HMW)(超过750kd)和低分子量(LMW)(87.5Kd)的谷氨酸脱羧酶(GAD)。动力学上,GAD活性的两个峰对L-谷氨酸表现出几乎相同的Km。如果浓缩步骤通过超滤进行,或者提取物在没有浓缩步骤的情况下进行层析,则仅出现LMW-GAD。如果在层析程序之前用Triton X-100处理GAD的乙醇提取物,HMW-GAD会从层析图谱中消失。先前分离的LMW-GAD的硫酸钠沉淀导致HMW-GAD峰重新出现。显然,HMW-GAD不是作为不同的分子实体存在;实际上,它可能是LMW-GAD的人为聚集物。