Effect of different nucleotides on oligomerization of Mcm4/6/7. The elution of Mcm4/6/7 from a 2.4 ml "mini" gel filtration column (Superose 6 PC 3.2/30; GE Healthcare) in the presence and absence of 5 mM adenine nucleotides is shown. A portion of the indicated fractions (above top panel) from each experiment was analyzed by GelCode Blue (Pierce) stained SDS-PAGE (6%). The migrations of Mcm4, Mcm6 and Mcm7 through the SDS gels are indicated to the right of each panel. Lanes containing molecular size markers are indicated (M) and their sizes are indicated on the left. The peak elution fractions of size standards (analyzed separately) are indicated below the bottom panel. The vertical lines mark the divisions between gels. In some experiments, nucleotide was added only to the protein (indicated by "load") and in some experiments nucleotide was added to both the protein and to the buffer used to equilibrate the gel filtration column ("load & column"). A) No nucleotide in either column buffer or load; B) 5 mM ATP in column buffer and load; C) 5 mM ATPγS added to load only; D) 5 mM ATP added to load only; E) 5 mM ADP in load only; and F) 5 mM ADP in column buffer and load. UV profiles are not shown since addition of nucleotide interfered with the signal from protein.
Ma et al. BMC Biochemistry 2010 11:37 doi:10.1186/1471-2091-11-37