摘要
目的 :探讨牙龈卟啉单胞菌FtsZ与FtsA之间相互作用的功能区域。方法 :通过缺失诱变法构建了C末端缺失变异型的PgFtsZ (ZΔC0 1,ZΔC0 2 ,ZΔC0 3) ,并表达和纯化了这些蛋白质 ,同时进一步通过免疫印迹法来鉴定这些C末端缺失变异型的PgFtsZ。然后 ,通过overlayassay法 ,以牛血清白蛋白为阴性对照 ,检测了这些C末端缺失变异型及野生型PgFtsZ和PgFtsA相互作用的特性。 结果 :野生型PgFtsZ与PgFtsA表现出极强的结合特性 ,然而去除C末端的 73个氨基酸残基的ZΔC0 1,则表现出极弱的结合特性 ,或者当进一步去除C末端的 12 8和 177个氨基酸残基的ZΔC0 2和ZΔC0 3,同ZΔC0 1一样也表现出与PgFtsA极弱的结合特性。 结论
Objective: To investigate the functional region of FtsZ-FtsA interaction in Porphyromonas gingivalis. Methods: Using deletion mutagenesis, a series of C-terminal deletion PgFtsZ mutants (viz., ZΔC01, ZΔC02, ZΔC03) were constructed, and expressed in Escherichia coli BL21(DE3)pLysS. These mutant proteins were purified by HiTrap SP column and identified by immunoblot method. FtsZ-FtsA interaction was measured by an overlay assay. Bovine serum albumin (BSA) was included as a negative control. Results: PgFtsA bound strongly to the wild-type PgFtsZ. However, PgFtsA was shown to be significantly decrease in the relative affinity of the interaction with the ZΔC01, a mutant form of PgFtsZ in which 73 amino acid residues were removed from the C-terminus. Furthermore, ZΔC02 and ZΔC03 (missing 128 and 177 amino acid residues from the C-terminus, respectively) were a similar extent the interaction as ZΔC01 did. Conclusion: The C-terminus of PgFtsZ plays a critical role in the FtsZ-FtsA interaction.
出处
《口腔医学研究》
CAS
CSCD
2004年第3期273-276,共4页
Journal of Oral Science Research
