Figure 1

The multiple domain structure of F-spondin and FS domain expression. (A) Schematic diagram of the domain structure of F-spondin showing the disulfide bond patterns of each domain and highly charged linkers between FS domain and TSR1, and between TSR5 and TSR6. (B) Size-exclusion chromatography of the recombinant FS domain. The recombinant protein was first purified using Ni-affinity chromatography. In the following FPLC runs with a Superdex™ 75 column (GE Healthcare Life Sciences), six protein standards were used to calibrate the column (data not shown). In the run of FS domain, the molecular mass of the principal peak was calculated to be 22.7 kDa, which was smaller than the calculated molecular mass of 27.23 kDa that does not include vector-derived sequences and N-linked glycans. SDS PAGE of the FS domain revealed a single band with a molecular mass greater than 30 kDa (insert).