"0"^^ . "Structure and dynamics of high and low affinity ATP binding sites of Na+/K+-ATPase"@en . . "N\u00EDzko- a vysokoafinn\u00ED m\u00EDsta pro ATP na Na+/K+-ATP\u00E1ze: jejich lokalizace, struktura a dynamika" . . . . "http://www.isvav.cz/projectDetail.do?rowId=GA204/98/0468"^^ . . . . "9"^^ . "9"^^ . . . . . "0"^^ . . . . . "1"^^ . "Na+/K+-ATP\u00E1za m\u00E1 dv\u011B ATP vazebn\u00E1 m\u00EDsta, kter\u00E1 b\u011Bhem katal\u00FDzy kooperuj\u00ED negativn\u011B na substr\u00E1ty ATP, pozitivn\u011B v\u0161ak s MANT-ATP. Vysokoafinn\u00ED m\u00EDsto pro ATP (E1ATP m\u00EDsto) m\u016F\u017Ee b\u00FDt modifikov\u00E1no fluorescein isothiokyan\u00E1tem (FITC) kovalentn\u011B v\u00E1zan\u00FDm na lysin 501. Z poznatk\u016F posledn\u00ED doby je zn\u00E1mo, \u017Ee E2ATP (n\u00EDzkofinn\u00ED m\u00EDsto) m\u016F\u017Ee b\u00FDt modifikov\u00E1no kovalentn\u011B v\u00E1zan\u00FDm erytrosinem isothiokyan\u00E1tem (ErITC). Zjistili jsme, \u017Ee na E2ATP vazebn\u00E9 m\u00EDsto se s vysokou afinitou v\u00E1\u017Ee tak\u00E9 DANS-8-N3-ATP. D\u00EDky sv\u011Bteln\u00E9 citlivosti vazby tohoto ATP analogu je mo\u017En\u00E9 E2ATP vazebn\u00E9 m\u00EDsto ozna\u010Dit dv\u011Bmi r\u016Fzn\u00FDmi fluorescen\u010Dn\u00EDmi zna\u010Dkami, co\u017E n\u00E1m, mimo jin\u00E9, umo\u017En\u00ED: 1) Lokalizovat aminokyselinov\u00E9 sekvence tvo\u0159\u00EDc\u00ED E2ATP vazebn\u00E9 m\u00EDsto. 2) Ur\u010Dit, pomoc\u00ED F\u00F6rsterova p\u0159enosu energie,vzd\u00E1lenost dvou ATP vazebn\u00FDch m\u00EDst. Odpov\u011B\u010F na tyto dv\u011B ot\u00E1zky je hlavn\u00EDm c\u00EDlem t\u00E9to pr\u00E1ce." . "Projekt p\u0159edstavuje \u0161pi\u010Dkov\u00FD v\u00FDzkum molekul\u00E1rn\u00ED biologie a biochemie. Z\u00EDskan\u00E9 v\u00FDsledky jsou v souladu s pl\u00E1ny grantov\u00E9ho projektu a zasahuj\u00ED do cel\u00E9 \u0159ady biologick\u00FDch obor\u016F. \u0158e\u0161en\u00ED grantu je zakon\u010Deno 12 publikacemi (10 ji\u017E vy\u0161lo, 2 p\u0159ijat\u00E9 do tisku). Fo"@cs . "Na+/K+-ATPase has two ATP sites which cooperate during catalysis in a negative coopartive way with ATP as substrate and in a positive cooperative way with MANT-ATP. The high affinity ATP site (E1ATP site) can be modified by fluorescein isothiocyanate (FITC) at lysine 501 in a covalent way. This lets the partial activity of the E2ATP site (K+-activated p-nitrophenylphosphatase unaffected. We recently became aware that the E2ATP site can be covalently modified by erythrosin isothiocyanata (EITC), sincethe K+-activated p-nitrophenylphosphatse is lost under these conditions. Moreover we became aware that the E2ATP site binds with high affinity the fluorescent DANS-8-N3-ATP. Since this ATP analog is photosensitive, it should be possible to label the E2ATsite by two different fluorescent labels opening up thereby the possibility to localize the amino acids forming the E2ATP site and to determine the distances between both ATP sites by F\u00F6rster energy transfer measurements."@en . "GA204/98/0468" .