Avtor/Urednik		Gregorc, Uroš
Naslov		Pomen cepitve izbranih predstavnikov na membranske proteine vezanih gvanilatnih kinaz s kaspazami med apoptozo
Tip		monografija
Kraj izdaje		Ljubljana
Založnik		Univerza v Ljubljani, Medicinska fakulteta
Leto izdaje		2005
Obseg		str. 101
Jezik		slo
Abstrakt		Apoptosis is a physiological form of cell death which ensures that the superfluous, damaged and potentially dangerous cells are safely removed from the organism. For the process to execute flawless a group of proteolytic enzymes, caspases, must be activated.Caspases involved in apoptosis can be divided in two subgroups. When cell receive death signal, initiator caspases are activated first. They primarily act as activators of executioner caspases, which in turn cleave various intracellular proteins thus causing the typical biochemical and morphological changes of apoptotic cells. One of the hallmarks of apoptosis is disruption of cell-cell contacts. Apoptotic cells detach from their neighbors causing no aftermath. Members of MAGUK (membrane associated guanylate kinase) proteins family are involved in establishing and maintaining cell-cell contacts. With their protein-protein interaction domains represent backbone to which actin cytoskeleton and transmembrane proteins, that physically connect neighbouring cells, are bound. Because of their connective function, multidomain structure and size, MAGUKS are possible caspase targets during apoptosis. We have selected three members of MAGUK family proteins, -DLG1, MAGI-1 and -3. First experiments were performed in vitro. cDNAs for selected MAGUKs were cloned in mammalian expression vector pcDNA3 and translated with TNT T7 Coupled Reticulocyte Lysate System (Promega). Proteins were labelled with 35S-Met for easy detection with radiography. Recombinant caspase-3, -6 and -7 were expressed in bacterial expression system and purified using affinity chromatography. They were checked for activity and used for cleavage of in vitro translated MAGUKs. (Abstract truncated at 2000 characters)
Deskriptorji		APOPTOSIS CYSTEINE PROTEINASES MEMBRANE PROTEINS PHOSPHOTRANSFERASES KERATINOCYTES CELLS, CULTURED CELL LINE, TRANSFORMED INTERCELLULAR JUNCTIONS GENETIC ENGINEERING FLOW CYTOMETRY RECOMBINANT PROTEINS MICROSCOPY, FLUORESCENCE