Submit Manuscript  

Article Details


Investigation of Folding of Purified Recombinant GRA1 Protein Using Web Based Protein Disorder Servers and Trypsin Digestion

[ Vol. 16 , Issue. 7 ]

Author(s):

Mert Doskaya, Ayse Caner, Aysu Degirmenci, Frances Jurnak and Yuksel Guruz   Pages 834 - 841 ( 8 )

Abstract:


The successful folding of a recombinant protein after expression and purification is essential for structural, biochemical and vaccination studies. Toxoplasma gondii recombinant GRA1 protein is a promising vaccine candidate against toxoplasmosis. In the present study, the folding of recombinant GRA1 protein has been evaluated by web based bioinformatics tools that predict protein folding. Subsequently, trypsin digestion, which is a simple indication of proper protein folding, has been used to determine whether recombinant GRA1 protein is likely to be folded. The results indicate that the recombinant GRA1 protein is predicted to be folded by most of the web based bioinformatics predictors. Moreover, in protease digestion experiments, the recombinant GRA1, which was purified to homogeneity without the use of denaturants, gives rise to a discrete band pattern that is indicative of a folded protein. Together, the results suggest that recombinant GRA1 protein is in a folded conformation, suitable for structural, biochemical and vaccination studies.

Keywords:

Toxoplasma gondii, dense granules, recombinant GRA1 protein, trypsin digestion, web based bioinformatics tools

Affiliation:

Department of Parasitology, Ege University Medical School, 35100 Turkey.



Read Full-Text article