Figure 4.
Figure 4. SUMO target sites are defined by their structural
context. (a) Position of four lysines in the N terminus of
E2-25K. (b) In the folded protein, Lys14 is the preferred
substrate. Wild type (WT) or indicated E2-25K mutant (500 ng), 1
g
SUMO1, 300 ng Aos1 -Uba2, 500 ng Ubc9 and ATP were incubated for
30 min at 30 °C. (c) In an unfolded peptide, Lys10 is preferred.
Indicated peptide (20 g)
1.5 g
SUMO-E1, 340 ng Ubc9, 15 g
SUMO and ATP were incubated for 0 -5 h. (d) In folded E2-25K
protein, Lys14 is preferred. Wild type or indicated F2-25K
mutant (7.5 g)
12 g
SUMO-E1, 425 ng Ubc9, 20 g
SUMO and ATP were incubated for 0 -2 h at 37 °C.
The above figure is reprinted
by permission from Macmillan Publishers Ltd:
Nat Struct Mol Biol
(2005,
12,
264-269)
copyright 2005.
g
SUMO1, 300 ng Aos1 -Uba2, 500 ng Ubc9 and ATP were incubated for
30 min at 30 °C. (c) In an unfolded peptide, Lys10 is preferred.
Indicated peptide (20