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Fig. 5 | Virology Journal

Fig. 5

From: Characterization of interactions between hepatitis C virus NS5B polymerase, annexin A2 and RNA – effects on NS5B catalysis and allosteric inhibition

Fig. 5

Analysis of the interplay between NS5B, AnxA2 and RNA determined by analysis of ternary complex formation using NS5B and the 8-mer RNA as analytes, and AnxA2 surfaces (3595 RU). The sensorgrams are aligned with respect to the baseline signal at the time for injection. a 1. Injection of 50 nM 8-mer RNA over an AnxA2 surface, providing a reference value for the RNA binding capacity of immobilized AnxA2. 2. 50 nM NS5B injected over the regenerated AnxA2 surface, resulting in a stable AnxA2-NS5B complex (as indicated by a stable new baseline after injection). 3. Injection of 50 nM 8-mer RNA over the AnxA2-NS5B surface. c 1. Injection of 50 nM NS5B over an AnxA2 surface, providing a reference value for the NS5B binding capacity of immobilized AnxA2. 2. 100 nM 8-mer RNA injected over the regenerated AnxA2 surface, creating a stable AnxA2-RNA complex. 3. 50 nM NS5B injected over the AnxA2-RNA complex surface.  b and d Schematic illustration of the experiments in  a and  c, respectively. e Schematic model of the interplay between NS5B and AnxA2 based on the presented data demonstrating formation of binary and ternary complexes, and where the 8-mer RNA interacts with AnxA2. Equilibrium 1 (E1) illustrates that AnxA2 can bind NS5B with strong affinity to form a stable binary complex. It has reduced NS5B polymerase activity and ability to interact with the allosteric inhibitor filibuvir. (E2) illustrates the interaction between AnxA2 and the 8-mer RNA, forming a binary complex that can interact with NS5B (E3), forming a ternary complex. This ternary complex can also be formed by the binding of the 8-mer RNA by the binary AnxA2-NS5B complex (E4). NS5B does not bind either the 8-mer RNA or the 31-mer RNA while in complex with AnxA2

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