Investigation of folding of purified recombinant GRA1 protein using web based protein disorder servers and trypsin digestion.
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.
Döşkaya
Mert
M
Department of Parasitology, Ege University Medical School, Bornova/Izmir 35100, Turkey. mert.doskaya@ege.edu.tr
Caner
Ayşe
A
Değirmenci
Aysu
A
Jurnak
Frances
F
Gürüz
Yüksel
Y
D000465:Algorithms; D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D019295:Computational Biology; D020407:Internet; D008969:Molecular Sequence Data; D017510:Protein Folding; D011994:Recombinant Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D013997:Time Factors; D014122:Toxoplasma / Q000737:chemistry* / Q000235:genetics; D014357:Trypsin / Q000378:metabolism*
D000953:Antigens, Protozoan; C412913:GRA1 antigen; D011994:Recombinant Proteins; D014357:Trypsin