Lammers J H, van Aalderen M, Peters A H, van Pelt A A, de Rooij D G, de Boer P, Offenberg H H, Dietrich A J, Heyting C
Department of Genetics, Agricultural University, Dreijenlaan 2, NL-6703 HA Wageningen, The Netherlands.
Chromosoma. 1995 Nov;104(3):154-63. doi: 10.1007/BF00352179.
The lateral elements (LEs) of synaptonemal complexes (SCs) of the rat contain major components with relative electrophoretic mobilities (Mr s) of 30000-33000, which are the products of a single gene. After one-dimensional separation of SC proteins on polyacrylamide-SDS gels, these components show up as two major bands, whereas upon two-dimensional electrophoresis they are resolved in at least 24 spots, which focus at pH 6.5 to 9.5. In this paper we show that these spots represent phosphorylation variants. For the analysis of the phosphorylation of the 30000- to 33000-Mr SC components during progression through meiotic prophase, we developed a procedure for isolation of fractions of testicular cells of the rat that are enriched in separate stages of meiotic prophase. Analysis of the 30000- to 33000-Mr SC components in these fractions by two-dimensional electrophoresis and immunoblotting showed that phosphorylated variants of the 30000- to 33000-Mr SC proteins occur throughout meiotic prophase. However, the extent of phosphorylation changes between early and mid-pachytene, when one phosphate group is probably added to each of the variants.
大鼠联会复合体(SCs)的侧生元件(LEs)包含相对电泳迁移率(Mr s)为30000 - 33000的主要成分,这些成分是单个基因的产物。在聚丙烯酰胺 - SDS凝胶上对SC蛋白进行一维分离后,这些成分显示为两条主要条带,而在二维电泳时,它们至少可分离为24个斑点,聚焦于pH 6.5至9.5。在本文中,我们表明这些斑点代表磷酸化变体。为了分析在减数分裂前期进程中30000至33000 Mr SC成分的磷酸化情况,我们开发了一种分离大鼠睾丸细胞组分的方法,这些组分在减数分裂前期的不同阶段富集。通过二维电泳和免疫印迹分析这些组分中30000至33000 Mr SC成分,结果表明30000至33000 Mr SC蛋白的磷酸化变体在整个减数分裂前期都存在。然而,在粗线期早期和中期之间磷酸化程度发生变化,此时每个变体可能添加了一个磷酸基团。