Wynik wyszukiwania w bazie Polska Bibliografia Lekarska GBL

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Tytuł oryginału: In situ activation of caspases and serine proteases during apoptosis detected by affinity labeling their enzyme active centers with fluorochrome-tagged inhibitors.
Autorzy: Grabarek Jerzy, Darzynkiewicz Zbigniew
Źródło: Exp. Hematol. 2002: 30 (9) s.982-989, il., bibliogr. 60 poz.
Sygnatura GBL: 305,749

Typ dokumentu:
  • praca opublikowana za granicą
  • tytuł obcojęzyczny

    Streszczenie angielskie: Activation of caspases is the key event of apoptosis. To detect this event in situ we applied fluorochrome-labeled inhibitors of caspases (FLICA) as affinity of activity centers of these enzymes. The FLICA are fluoroscein- or sulforhodamine-conjugated peptide-fluoromethyl ketones that covalently, with 1:1 stoichiometry, bind to enzymatic centers of caspases; the specifity is provided by the peptide sequence of amino acids. Similarly, we applied fluorescent inhibitors of serine proteases (FLISP) to detected activite sites of the latter enzymes. Exposure of live cells to FLICA of FLISP led to uptake of these ligands and their binding to activated caspases or active sites og serine proteases; the unbound reagents were removed by cell rinse. Only cells undergoing apoptosis were labeled with FLISP or FLICA. Intracellular binding sites of FLICA are consistent with known localization of caspases. Covalent binding of FLICA or FLIPPS allowed us to identify the labeled proteins by immunoblotting: the proteins that bound individual FLICAs had molecular weight between 17 and 22 kDa, which corresponds to large subunits of the caspases; two proteins reacting with FLISP were about 57 and 60 kDa, which suggests that they are novel enzymes. Detection of caspases or serine proteases activation can be combined with other markers of apoptosis or cell cycle for multiparametric analysis by flow or laser ascanning cytometry. Being caspases inhibitors, FLICA arrest the process of apoptosis and prevent cell distintegration. The stathmo-apoptotic assay was developed...

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    Tytuł oryginału: Zastosowanie inhibitorów proteaz znakowanych fluorochromami do cytometrycznej oceny aktywacji proteaz cysteinowych i serynowych w czasie apoptozy : praca doktorska
    Autorzy: Grabarek Jerzy, Urasińska Elżbieta (promot.).; Akademia Medyczna [w Szczecinie], [Medical College Brander Cancer Research Institute in New York]
    Źródło: 2002, 50 k. : il., tab., bibliogr. 113 poz., streszcz., maszyn.
    Sygnatura GBL: 45/20297

    Typ dokumentu:
  • praca doświadczalna
  • praca doktorska

    Wskaźnik treści:
  • in vitro

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