Oligomerization of sponge NmeGp1Sd protein and human Nme1 as control. (A) Cross-linking NmeGp1Sd (S1) and Nme1 (H1) with glutaraldehyde. Purified recombinant protein NmeGp1Sd (8 μg) was pre-incubated in PBS at room temperature with 25 mM of glutaraldehyde to initiate the cross-linking. After quenching, the reaction product was subjected to 12.5% SDS-PAGE gel followed by staining with Coomassie brilliant blue. (B) Size exclusion chromatography. Recombinant human Nme1 and sponge NmeGp1Sd were loaded onto Bio-Sil SEC 250 gel-filtration columns (300 mm × 7.8 mm) and eluted with Nm23 buffer at a flow rate of 0.5 mL/min. Black line represents human Nme1 and grey line sponge NmeGp1Sd protein.
Perina et al. BMC Evolutionary Biology 2011 11:87 doi:10.1186/1471-2148-11-87