Hartmann H, Noegel A A, Eckerskorn C, Rapp S, Schleicher M
Max-Planck-Institut für Biochemie, Martinsried, Federal Republic of Germany.
J Biol Chem. 1989 Jul 25;264(21):12639-47.
Polyclonal and monoclonal antibodies against purified 32/34-kDa F-actin-capping protein from Dictyostelium discoideum were generated and used to isolate clones coding for both subunits from a lambda gt11 expression library. In addition genomic clones were isolated for the 34-kDa subunit. The sequences of the corresponding inserts were determined and the amino acid sequences of the proteins deduced. The amino acid sequences have been confirmed by sequencing tryptic peptides of both subunits. As judged from immunological data and sequence comparison, the subunits are completely different. Each of them is encoded by a single copy gene. The mRNAs are present throughout all stages of development. Severing proteins, which exert also capping activity, show no sequence similarity with the cap 32/34-kDa protein. However, a pronounced homology between the 32-kDa subunit and a hypothetical yeast protein was found indicating the presence of a protein homologous to the 32-kDa subunit of the D. discoideum capping protein in yeast.
制备了针对盘基网柄菌纯化的32/34-kDa F-肌动蛋白封端蛋白的多克隆和单克隆抗体,并用于从λgt11表达文库中分离编码两个亚基的克隆。此外,还分离了34-kDa亚基的基因组克隆。确定了相应插入片段的序列,并推导了蛋白质的氨基酸序列。通过对两个亚基的胰蛋白酶肽段测序,证实了氨基酸序列。从免疫学数据和序列比较判断,这两个亚基完全不同。它们各自由一个单拷贝基因编码。这些mRNA在发育的所有阶段都存在。同样具有封端活性的切割蛋白与32/34-kDa封端蛋白没有序列相似性。然而,发现32-kDa亚基与一种假定的酵母蛋白之间存在明显的同源性,这表明酵母中存在与盘基网柄菌封端蛋白32-kDa亚基同源的蛋白。