Figure 1.Effects of phosphorylation and Pin1 binding on p66Shc.(a-c) The
p66CH2CB-dependent ROS-generation is enhanced by phosphorylation of Ser36
but inhibited in the presence of Pin1. Changes in fluorescence of 10 μM H2DFFDA
were recorded after addition of 20 μM p66CH2CB WT (a) or the
p66CH2CB-Ser36Asp mutant simulating Ser36 phosphorylation (a-c),
followed by 85 μM Na-dithionite (DN) and 50 μM CuSO4 (Cu) in the
presence (b+c) or absence of Pin1 (a-c) and/or the dipeptide
Ala-Pro (c). (d) p66CH2CB-induced mitochondrial rupture is
inhibited by phosphorylation of Ser36. Mitochondrial rupture was induced
after addition of 7 μM CaCl2 by addition of 20 μM p66CH2CB WT or
Ser36Asp and monitored photometrically. The initial sensitization with CaCl2
was omitted for clarity. (e) H2O2 oxidizes
p66CH2CB. 10 μg p66CH2CB were incubated with 0.005 % H2O2
and subjected to non-reducing SDS-PAGE
