Figure 4: Unfolding and degradation of stable proteins by hexYME1L.
From: Engineered AAA+ proteases reveal principles of proteolysis at the mitochondrial inner membrane
(a) SDS–PAGE showing degradation of β20-λcI-N (20 μM) by hexYME1L (1 μM) in the presence (+) and absence (−) of ATP and a plot of the loss of β20-λcI-N over time. (b) SDS–PAGE showing degradation of circularly-permuted variants of GFP (20 μM) by hexYME1L (1 μM) in the presence (+) or absence (−) of ATP. (c) Loss of GFP variants over time from degradation experiments in c. (d) Rate of cp7-SFGFP-β20 degradation against increasing substrate concentration displaying a nonlinear least-squares fitting to the Michaelis–Menten equation (kdeg=0.11 GFPs min−1 enz6−1; KM=7.1 μM). **P≤0.01, ***P≤0.001, ****P≤0.0001 calculated from an unpaired Student’s t-test (two-tailed). All data shown are from independent experiments and error bars indicate±s.e.m. (n=3).
