Mouse LSECtin as a model for a human Ebola virus receptor

Pipirou, Zoi; Powlesland, Alex S; Steffen, Imke; Pöhlmann, Stefan; Taylor, Maureen E; Drickamer, Kurt

The CRD from mouse LSECtin was initially expressed in a bacterial system previously used for production of the human protein (Powlesland et al. 2008) , in which the portion of the cDNA encoding the CRD is inserted immediately after the bacterial ompA signal sequence. The signal sequence allows folding of the CRD in the periplasm, so the protein could be extracted and purified directly by affinity chromatography on immobilized sugars (data not shown). However, for the mouse CRD, more efficient expression was obtained by production of the CRD as inclusion bodies that were dissolved in guanidine hydrochloride and renatured by dilution and dialysis ( Figure 1C ). Similar yields were obtained by purification on either fucose-or mannose-Sepharose. However, due to the relatively weak affinity of individual CRDs for the monosaccharide ligands, a 10-mL column was required to achieve effective purification. The CRD was used directly in the solidphase binding assays.