Kübler D, Pyerin W, Kinzel V
Eur J Cell Biol. 1982 Feb;26(2):306-9.
The ability of phosvitin and histone to serve as substrates for possible protein kinase(s) at the cell surface was examined with regard to their suitability as indicators of such activity. While phosvitin did not interfere with the membrane barrier of HeLa cells, 3T3 and SV 3T3 cells, histone caused severe damage as indicated by both uptake of viability stains Trypan Blue and diamidino-phenylindol and by release of intracellular compounds such as lactate dehydrogenase, cAMP-dependent protein kinase(s), and metabolically prelabelled proteins. Where intactness of the cell membrane is prerequisite for verification of ecto-protein kinase, histone cannot be used as substrate. In contrast, we found that phosvitin is suitable for assays of cell surface located protein kinase activity.
就磷蛋白和组蛋白作为细胞表面可能的蛋白激酶底物的能力而言，研究了它们作为此类活性指标的适用性。虽然磷蛋白不会干扰HeLa细胞、3T3细胞和SV 3T3细胞的膜屏障，但组蛋白会造成严重损伤，这可通过活力染料台盼蓝和双脒基苯基吲哚的摄取以及细胞内化合物如乳酸脱氢酶、cAMP依赖性蛋白激酶和代谢预标记蛋白的释放来表明。细胞膜的完整性是验证胞外蛋白激酶的前提条件，因此组蛋白不能用作底物。相比之下，我们发现磷蛋白适用于检测细胞表面的蛋白激酶活性。
