Lewis Tamra E, Milam Terry D, Klingler Douglas W, Rao Prema S, Jaggi Meena, Smith David J, Hemstreet George P, Balaji K C
Division of Urological Surgery, University of Nebraska Medical Center, Omaha, NE 68198-2360, USA.
Urol Oncol. 2005 Nov-Dec;23(6):407-12. doi: 10.1016/j.urolonc.2005.04.002.
We have previously described that tissue transglutaminase (tTG) is a high level phenotypic biomarker in prostate cancer, which is down regulated in prostate cancer and surrounding premalignant field compared to benign prostate glands. To understand the function of tTG in prostate cancer, we sought to identify proteins that interact with the transglutaminase moiety of tTG using a human prostate cancer complementary deoxyribonucleic acid library in a Yeast 2-Hybrid system. The Yeast 2-Hybrid experiments identified a strong and novel interaction between the transglutaminase moiety and protein kinase A anchor protein 13 (AKAP13), which was quantified by beta-galactosidase assay, confirmed in vitro by immunoprecipitation experiments using PC3 prostate cancer cell lysates, and in vivo colocalization was confirmed by immunofluorescence studies in PC3 cells. Because AKAP plays a major role in protein kinase A and Rho protein mediated signaling, functional studies are underway to elucidate the significance of tTG-AKAP13 interaction in prostate cancer.
我们之前曾描述过，组织转谷氨酰胺酶（tTG）是前列腺癌中的一种高水平表型生物标志物，与良性前列腺腺体相比，其在前列腺癌及周围癌前区域中表达下调。为了解tTG在前列腺癌中的功能，我们试图在酵母双杂交系统中利用人前列腺癌互补脱氧核糖核酸文库来鉴定与tTG转谷氨酰胺酶部分相互作用的蛋白质。酵母双杂交实验确定了转谷氨酰胺酶部分与蛋白激酶A锚定蛋白13（AKAP13）之间存在强烈且新颖的相互作用，通过β-半乳糖苷酶测定对其进行了定量分析，利用PC3前列腺癌细胞裂解物进行免疫沉淀实验在体外对其进行了证实，并且通过对PC3细胞进行免疫荧光研究在体内证实了共定位。由于AKAP在蛋白激酶A和Rho蛋白介导的信号传导中起主要作用，目前正在进行功能研究以阐明tTG-AKAP13相互作用在前列腺癌中的意义。
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