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. 2016 Feb 19;291(16):8686–8700. doi: 10.1074/jbc.M115.713628

FIGURE 1.

FIGURE 1.

Expression and purification of CTR ECD and RAMP1/2-CTR ECD fusion proteins in HEK293T cells. A, protein constructs designed for this study. Five pairs of (Gly-Ser) were used as a linker between RAMP1/2 and CTR ECDs. Amino acid numbers used for the ECDs are indicated above the diagram. The schematic shown is not scaled with real protein size. B, gel filtration elution profiles of MBP-CTR ECD, MBP-RAMP1-CTR ECD, and MBP-RAMP2-CTR ECD fusion proteins. C, left image is non-reducing SDS-PAGE analysis. Molecular mass markers are shown in the 1st lane and are labeled in kDa. Tris-glycine native gel analysis is shown in the right panel where a slight upward shift was observed for the E101A mutant protein due to the loss of a negative charge on Glu-101. The gels were stained with Coomassie Brilliant Blue.