Research Paper Volume 1, Issue 2 pp 254—265
Identification of Peroxiredoxin 1 as a novel interaction partner for the lifespan regulator protein p66Shc
Figure 2. Identification of the novel p66Shc interaction partner Prx1. (a) In vivo pull down and subsequent MudPIT analyses identified Prx1 as an interaction partner of p66CH2CB. The proteins discussed in the text (Prx1, Prx3 and Hsp70) are highlighted in grey. (b-d) p66CH2CB interacts with dimeric and decameric Prx1 in a 1:1 stoichiometry and perturbs its decameric arrangement. 20 μg p66CH2CB WT (b) or p66CH2CB-Cys59Ser (c) were incubated with 40 μg Prx1 WT or 20 μg Prx1-Cys83Ser, respectively, for 1 hour at room temperature and subjected to BN-PAGE (b+c). Lanes 1 and 2 from (b) are repeated in (c) for clarity. The p66CH2CB-Cys59Ser/Prx1-Cys83Ser interaction complex is marked by an arrow. (d) Bands labelled 1-7 in panel (b) were subjected to a second separation dimension, denaturing SDS-PAGE.