Publication: Sığır plazmasındaki K vitaminine bağımlı proteinlerin iki boyutlu elektroforezle ayrılması
Abstract
1. ÖZET Protrombin, faktör VII, faktor IX, faktör X, protein C, protein S ve protein Z K vitaminine bağlı proteinler (Gla proteinleri) olarak bilinir ve kromotogrofik metotlarla sistematik olarak saflaştırılmışlardır. Hem klinik çalışma hem de araştırma amacıyla, daha kısa sürede ve eş zamanlı olarak K vitaminine bağımlı proteinleri plazmadan izole etmek/ miktar tayini yapmak yararlı olabilir. Bu çalışmanın amacı 2-D elektroforeziyle Gla proteinlerini ayırmaktır. Baryum sitrat adsorpsiyonu ve amonyum sülfat elüsyonuyla normal sığır plazmasından izole edilen Gla proteinleri, 2-D elektroforezine tabi tutuldu. Birinci boyut bir pH gradientinde izoelektrik odaklama ve ikinci boyut SDS elektroforeziydi. Coomassie Blue R-250 ile boyanan jellerde görülen 8 den fazla protein spotu, iyi bilinen plazma Gla protein sayısından daha fazlaydı. Bunların bazıları, yıkım ürünleri veya plazma proteinlerinin kontaminasyonu sebebiyle olabilir. 2-D ile plazmadaki K vitaminine bağımlı proteinlerin ayrılması mümkün olmuştur. 2-D örneklerinin ileride kütle spektrometresiyle değerlendirmesi araştırma/ klinik çalışmalar için daha faydalı bilgi sağlayacaktır. Insan plazmasından elde edilen K vitaminine bağımlı proteinlere ait 2-D örneklerinin, bilgisayar aracılı dansiyometrik eşzamanlı incelenmesi/ miktar tayini klinik çalışmalarda faydalı olabilir. 2.
RESOLUTION OF BOVINE PLASMA VITAMIN K DEPENDENT PROTEINS BY TWO DIMENSIONAL GEL ELECTROPHORESIS Prothrombin, factor VII, factor IX, factor X, protein C, protein S and protein Z are well-known vitamin K dependent (Gla proteins) proteins which have been purified systematically by time consuming chromatographic methods. It may be useful in clinical practice/ research to perform a less time consuming procedure for simultaneous isolation/ quantification of Gla proteins from plasma. The purpose of this study is to resolve the Gla proteins by two-dimensional (2-D) gel electrophoresis. Gla proteins isolated from normal bovine plasma by barium citrate adsorption and ammonium sulfate elution was subjected to 2-D gel electrophoresis. It was carried out with a pH gradient in the first dimension and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the second dimension. More than 8 protein spots were visualized by staining with Coomassie Blue R-250. The number of protein spots was more than the number of well-known plasma Gla proteins. Some of them might be contamination of plasma proteins or degradation products. 2-D method can be used for resolution of human plasma Gla proteins. Further mass spectrophotometric evaluation of the 2-D pattern will be providing useful data for research/ clinical studies. Computer assisted densitometry of the human protein pattern of 2-D may be helpful in clinical studies for simultaneous quantification of plasma Gla proteins.
RESOLUTION OF BOVINE PLASMA VITAMIN K DEPENDENT PROTEINS BY TWO DIMENSIONAL GEL ELECTROPHORESIS Prothrombin, factor VII, factor IX, factor X, protein C, protein S and protein Z are well-known vitamin K dependent (Gla proteins) proteins which have been purified systematically by time consuming chromatographic methods. It may be useful in clinical practice/ research to perform a less time consuming procedure for simultaneous isolation/ quantification of Gla proteins from plasma. The purpose of this study is to resolve the Gla proteins by two-dimensional (2-D) gel electrophoresis. Gla proteins isolated from normal bovine plasma by barium citrate adsorption and ammonium sulfate elution was subjected to 2-D gel electrophoresis. It was carried out with a pH gradient in the first dimension and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the second dimension. More than 8 protein spots were visualized by staining with Coomassie Blue R-250. The number of protein spots was more than the number of well-known plasma Gla proteins. Some of them might be contamination of plasma proteins or degradation products. 2-D method can be used for resolution of human plasma Gla proteins. Further mass spectrophotometric evaluation of the 2-D pattern will be providing useful data for research/ clinical studies. Computer assisted densitometry of the human protein pattern of 2-D may be helpful in clinical studies for simultaneous quantification of plasma Gla proteins.