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. 2013 Apr 2;110(16):6394–6399. doi: 10.1073/pnas.1300480110

Fig. 3.

Fig. 3.

Localization of interacting residues. (A) Percent conversion of CPIII-Long [340 nM; both WT and the K18A, K35Q, and K45Q mutants] by BMP-1 (16 nM) in the absence and presence of PCPE-1 (340 nM), incubated for 1 h at 37 °C. Analysis by SDS/PAGE (reducing conditions) and Sypro Ruby staining, followed by fluorescence quantitation. Error bars correspond to SDs (n = 3). (B) CD spectra of WT CPIII-Long and its K18A, K35Q, and K45Q mutants, all produced in HEK 293T cells, analyzed at 25 °C in 20 mM Tris·HCl (pH 7.4), 2.5 mM CaCl2. Also shown is the CD spectrum for CPIII-Long produced in insect cells (WT-Bac), which is indistinguishable from that for protein produced in 293T cells. Protein concentrations 219–400 µg/mL. (C) Percent conversion of CPIII-Long (350 nM) by BMP-1 (16 nM) in the absence and presence of full-length PCPE-1 or its CUB1CUB2 fragment [both WT (C1C2) and the D191A mutant], all at 350 nM, incubated for 1 h at 37 °C. Analysis by SDS/PAGE (reducing conditions) and Sypro Ruby staining, then fluorescence quantitation. Error bars correspond to SDs (n = 4).