Fig. 2: Amino acid sequence of VHH12G2 and its binding characteristics for Ube2G2. | Communications Biology

Fig. 2: Amino acid sequence of VHH12G2 and its binding characteristics for Ube2G2.

From: A nanobody that binds to the backside of the ubiquitin conjugating enzyme Ube2G2 differentially affects interactions with its partner E3 Ligases

Fig. 2

A Amino acid sequence of VHH12G2 with complementarity determining regions (CDRs) highlighted according to the IMGT database56. B Size exclusion chromatography (SEC) profiles that illustrate binding of Ube2G2 to VHH12G2 with milli absorbance units (mAU) plotted against fraction numbers. The elution profiles for Ube2G2 (green), VHH12G2 (blue) and a 2:1 molar ratio mixture of VHH12G2: Ube2G2 (red) are shown. Protein fractions from the indicated peaks were analyzed by SDS-PAGE (shown on the right). Note the co-elution of VHH12G2 and Ube2G2, distinguishable by a subtle difference in migration, in the peak fraction (red trace). C Microscale thermophoresis of the affinity (Kd) measurement of VHH12G2 for Ube2G2 tagged with Alexa Fluor 647. The binding data, averaged from three independent experiments, are presented with the estimated binding constant indicated. The raw relative fluorescence data for all MST runs is displayed on the right.

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