摘要
应用Sephacryl S-100-HR凝胶层析、高效液相色谱(HPLC)和反相高效液相色谱(R-HPLC)分析,从巨噬细胞条件培养基中纯化了巨噬细胞源性神经营养因子(MΦDNF)。该因子在SDS-聚丙烯酰胺凝胶电泳上呈现一条单一蛋白带,其分子量为60.5kD,等电点约5.1。其氨基酸组成含有较高比例的天冬氨酸、谷氨酸、亮氨酸和赖氨酸。纯化的MΦDNF能支持体外培养的小脑皮质神经元的存活,增强其活性及促进神经元突起的生长。其最大效应浓度为500—1000ng/ml。
Macrophage-derived neurotrophic factor (MΦDNF) is purified from macrophage conditioned medium by a procedure consisting of column chromatography with Sephacryl S-100-HR, high-performance liquid chromatography (HPLC),and a final step using reverse-phase HPLC. The product shows a single protein band in sodium dodecyl sulfate-polyacrylamide gel. It
has a molecular weight of 60. 5kD and an isoelec-tric point of pI 5. 1 and contains more leucine, ly-sine, glutamine and aspartic acids in its amino acid composition. Purified MΦDNF can promote the survival, activity, and neurite outgrowth of cultured cerebellar cortical neurons and that this effect reachs maximal levels with concentrations of the MΦDNF ranging from 500-1000ng/ml.
出处
《实验生物学报》
CSCD
1998年第2期137-145,共9页
Acta Biologiae Experimentalis Sinica
基金
国家自然科学基金(批准号39470356)
国家教委博士点科研基金(批准号96056909)~~
关键词
巨噬细胞源性
神经营养因子
纯化
鉴定
Macrophage. Macrophage-derived neurotrophic factor. High-performance liquid chro-matography. Sephacryl S-100-HR gel chromatography. Automated colorimetric mi-croassay.
