Příprava a charakterisace rekombinantního dermcidinu jako potenciálního proteinového partnera glutamátkarboxypeptidasy II

Tužil, Jan

A process of forming new blood vessels is necessary for tumour viability and expansion. Without vasculature, tumour stops growing at a size of millimeters. Some tumours, however, undergo an angiogenic switch and start to build up their own vascular architecture. The rate of apoptosis then decreases and the tumour becomes invasive. There are many factors that control the process of physiological angiogenesis. These might or might not relate to tumour tissue as well. Glutamate carboxypeptidase II (GCPII; EC 3.4.17.21) is a type II transmembrane glycoprotein with two known enzymatic activities. GCPII expression is upregulated in prostate cancer and also highly expressed in tumour-associated neovasculature even though none of these enzymatic functions was observed on the endothelium. Although numerous researches suggested that GCPII might serve as a receptor, no natural ligand has been identified yet. Preliminary experiments performed in our laboratory indicated some proteins to be possible natural ligands of GCPII. Therefore, we chose one of them- dermcidin, cloned and expressed this protein in mammalian cells. We investigated its possible interaction with GCPII introducing new detection system utilizing FLAG-tag however, we were not able to approve neither disapprove its interaction in vitro.

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