Figure 4
From: Crystal structure of the DNA-binding domain of Myelin-gene Regulatory Factor
MyRF DBD mutation disturbs trimer formation but does not affect auto-cleavage of MyRF. (A) The auto-cleavage of MyRF and its mutants were analyzed by 12% SDS-PAGE. M: protein marker. (B) Elution profiles from gel filtration chromatography of MyRF DBD and its mutant. Calibration curve of proteins with known molecular weights (ferritin (443 kDa), bovine serum albumin (BSA; 66 kDa), chicken ovalbumin (44kD); Sigma-Aldrich) was shown. The molecular weight of these proteins was plotted against their calculated Ve/Vo and fitted by exponential regression analysis. On the basis of the calculation, the Ve/Vo of wild type MyRF DBD results in a molecular weight of 110 kDa, and the Ve/Vo of its mutant results in a molecular weight of 35 kDa, indicating the trimeric and the monomeric form of protein respectively. The Ve/Vo of wild type MyRF DBD core only (after treated by limited proteolysis of trypsin) results in a molecular weight of 70 kDa, indicating the trimeric form too.
