| Summary: | 碩士 === 國立中正大學 === 分子生物研究所 === 96 === Polo-like kinase 1 (Plk1) is a serine-threonine protein kinase implicated in the regulation of multiple aspects of mitosis. Plk1 contains a kinase domain in the amino-terminal region and a common structure motif called polo box domain in the carboxyl-terminal region. It was found that Plk1 is overexpressed in many types of cancer. To further our studies in the functions of Plk1, we raised and purified an anti-Plk1 antibody using the Plk1 C-termainal linker-PBD as the immunogen. The antibody was tested successfully in western blotting, in immunofluorscence staining, and in immunohistochemistry. We used this antibody to examine the expression level of Plk1 in the biopsies from the patients inflicted with the head and neck cancer and found that high levels of Plk1 were found in the advanced cancer. In the 48 samples, Plk1 is indeed overexpressed in the head and neck cancer and that the expression levels appear to be associated with advanced stages.
Additionally, phosphorylation of Plk1 is an important topic. Since, it has to be phosphorylated to enhance the activity of Plk1. We designed a series of experiments. We compared the different phosphorylation status of two purified linker-PBD T498E and T498V proteins. In pull-down assay and two dimensional electrophoresis,it is five proteins affected by phosphorylation at least. Totally, we identified three proteins as hnRNP K、β-tubulin and actin by LC MS/MS.
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