DeArmond S J, Lee Y L, Kretzschmar H A, Eng L F
J Neurochem. 1986 Dec;47(6):1749-53. doi: 10.1111/j.1471-4159.1986.tb13084.x.
Twenty-day-old mice received a single tail vein injection of [guanido-14C]arginine. The cytoskeleton was extracted from the spinal cords at varying lengths of time thereafter. Glial fibrillary acidic protein (GFAP) formed a distinct, broad band that was widely separated from other protein bands in one-dimensional polyacrylamide gels. The purity of the GFAP band was verified by Western blot analysis of one- and two-dimensional electrophoretic patterns. In addition, enzyme-linked immunosorbent assay and quantitative Western blot analysis indicated that 95% of the total spinal cord GFAP was extracted in the cytoskeletal preparation. The specific activity of GFAP was obtained by eluting the protein from the cytoskeletal GFAP band in preparative one-dimensional gels. Specific activity reached a peak 2 h after injection with [14C]arginine. Forty percent of the incorporated radioactivity was still present in cytoskeletal GFAP at 9 weeks, indicating that a significant proportion of glial filaments turns over relatively slowly in vivo.
20日龄的小鼠接受了一次尾静脉注射[胍基-14C]精氨酸。此后在不同时间从脊髓中提取细胞骨架。在一维聚丙烯酰胺凝胶中，胶质纤维酸性蛋白（GFAP）形成了一条明显的宽带，与其他蛋白带广泛分离。通过对一维和二维电泳图谱进行蛋白质印迹分析，验证了GFAP带的纯度。此外，酶联免疫吸附测定和定量蛋白质印迹分析表明，在细胞骨架制备物中提取了脊髓中95%的总GFAP。通过在制备性一维凝胶中从细胞骨架GFAP带洗脱蛋白质来获得GFAP的比活性。注射[14C]精氨酸后2小时，比活性达到峰值。9周时，40%的掺入放射性仍存在于细胞骨架GFAP中，这表明相当一部分胶质丝在体内的周转相对较慢。
