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- W2238555732 abstract "Miyeloperoksidaz (MPO) enzimi, lokoferez ile elde edilen insan lokositlerinden saflastirildiktan sonra kinetik ozellikleri belirlendi; MPO tarafindan katalizlenen nitrit oksidasyonu ve tirozin nitrasyonu arastirildi. Serum fizyolojik ile yikanan lokositlerden MPO enzimi %1 (w/v) HETAB iceren fosfat tamponu icinde cozunurlestirildi. Cozunurlestirilmis enzim fraksiyonu, Concanavalin A-Sepharose kromatografisi ve pH:8’de yapilan CM-Sephadex iyon-degistirici kromatografisi ile % 70.3 verimle saflastirildi. Saflastirilan enzim indirgen kosullardaki SDS-PAGE’de 57000 ve 13100 Dalton molekul agirliginda iki protein bandi verdi. Enzimin spektral analizinde, saflastirilan MPO’nun Reinheit Zahl (RZ) degeri 0.86 olarak bulundu. Bu veri enzimin en az % 99 oraninda saf oldugunu gosterdi. Substrat olarak H2O2 ve tetrametilbenzidin kullanilarak saflastirilan enzimin bu iki substrati icin kinetik ozellikleri belirlendi. MPO’nun H2O2 icin Km ve Vmax degerleri 0.727mM ile 273±8 IU/mg protein; tetrametilbenzidin icin ise sirasi ile 0.111 mM ve 283.04±39 IU/mg protein olarak bulundu. MPO, fagozom-ici pH degerleri civarinda (pH 4.5-6) ve H2O2 varliginda, nitriti substrat olarak kullanarak oksidasyonunu katalizlemektedir. Daha dusuk pH’da nitritin oksidasyonu MPO’dan bagimsizdir. pH:7 civarinda ise MPO nitrit oksidasyonuna (aktivasyonuna) etkisizdir. Nitritin oksidasyonu zamana bagli olup, MPO tarafindan kullanilan nitritin yaklasik % 50’si nitrata cevrilmektedir. MPO, nitrit ve H2O2 varliginda, serbest tirozinlerin veya proteine bagli tirozinlerin nitrasyonunu katalizleyebilir. Saflastirilan MPO’nun fagozom ici pH araliklarinda (pH:5-6), etkili olarak tirozin nitrasyonunu katalizledigi; nitrasyon hizinin substratlari olan nitrit, H2O2 ve tirozin derisimlerine ayri ayri bagli oldugu goruldu. MPO tarafindan katalizlenen tirozin nitrasyonunun, iki onemli fizyolojik elektron verici antioksidan bilesik olan askorbik asit ile indirgenmis glutatyon tarafindan karisik-tip inhibisyon mekanizmasi ile inhibe edildigi bulundu." @default.
- W2238555732 created "2016-06-24" @default.
- W2238555732 creator A5044118497 @default.
- W2238555732 date "2013-01-01" @default.
- W2238555732 modified "2023-09-23" @default.
- W2238555732 title "Lökosit Miyeloperoksidaz Enziminin Saflaştırılması, Kinetik Özellikleri ile Reaktif Türlerin Oluşumundaki Etkilerinin Araştırılması" @default.
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