KAx-3和AK127细胞发育期间的骨架蛋白组分比较研究

Comparative Investigation of Cytoskeleton Proteins between KAx-3 and AK127 Cells during Dictyostelium Development

采用生化抽提骨架蛋白和 SDS-聚丙烯酰胺凝胶电泳等方法比较盘基网柄菌 AK127细胞和 KAx-3细胞骨架蛋白的结果显示,各发育时期的 KAx-3和 AK127细胞骨架蛋白在含量和组分方面存在一定的差异．在整个发育阶段两种类型细胞共有的蛋白组分是103．4 kD、49．6 kD、 44．2 kD、30．8 kD、19．8 kD 和13．5 kD 条带,其中44．2 kD 和30．8 kD 是最稳定的细胞骨架成分,并不因 AK127细胞缺失 gp150分子而有所改变．它们与其他骨架蛋白组分一起为细胞提供有力的支持,保证发育的顺利进行．差别最为明显的蛋白条带是87．2 kD、68．2 kD 和40．7 kD 蛋白,由于同时期的突变型细胞不能显示这些蛋白条带；说明这些条带是发育所需的蛋白．值得注意的是突变型细胞也出现一条特征性的21．1 kD 的蛋白条带．分析认为这些变化一方面与盘基网柄菌发育过程有关,另一方面与突变细胞缺乏 gp150分子有密切关系．

Cells of Dictyostelium discoideum are trigged to undergo development when they are depleted their food source. Cell cytoskeleton proteins play an important role for completing the development of Dictyostelium discoideum. To study the change of cytoskeleton proteins during Dictyostelium discoideum development, we used the wild-type cells KAx-3 and mutant cells AK127; cytoskeleton ghost have been prepared by biochemical extraction with TrotonX-100, cytoskeleton proteins were separated using SDS-PAGE and scan by BIO-PROFILE of VILBER LOURMAT. The datum showed that the 103.4 kD,49.6 kD,44. 2 kD,30.8 kD,19.8 kD and 13.5 kD were shared by KAx-3 and AK127 cells; in that 44.2 kD and 30. 8 kD were the most stable. They may take some support for cell development with other cytoskeleton proteins. The most different existed in some proteins, for example 87.2 kD, 68.2 kD and 40.7 kD, those proteins can not be expressed in the mutant cells at the same developmental stage of wild type cells. This result indicates those proteins are required for Dictyostelium cells development. It is noteworthy that mutant cell showed the characteristic 21.1 kD proteins after 12 h development. These obvious difference related to the process of development, in other hand, also related to the mutant cell lacking gp150 protein.