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Figure 2.
Figure 2 Crystal structure of the Nup50:importin-  complex
and competition assays using structure-based mutants showed that
Nup50 binds at two sites on importin- ,
and the NLS displacement by Nup50 requires interaction with both
sites on importin- .
(A) Overview of the Nup50:importin- complex
with the 2F[o]-F[c] map around Nup50 contoured at 1  superimposed.
Nup50 residues are shown in ball-and-stick format. (B)
Electrostatic potential on importin- ,
with Nup50 removed, shaded from -9 kT/e (red) to +9 kT/e (blue)
calculated using GRASP (Nicholls et al, 1991). (C) FRET-based
competition assay. Emission profiles of 0.2  M
BFP-  IBB
importin- and
0.2 M
SV40 NLS-GFP without (red) 1 M
Nup50 (residues 1-109) or with (black) 1 M
Nup50 (residues 1-109, wild type) or with (blue) 1 M
Nup50 (residues 1-109, K3E/R4D), or with (green) 1 M
Nup50 (residues 1-109, R38A/R45D). (D) FRET-based competition
assay. Emission profiles of 0.2 M
BFP- IBB
importin- and
0.2 M
NP NLS-GFP without (red) 1 M
Nup50 (residues 1-109) or with (black) 1 M
Nup50 (residues 1-109, wild type) or with (blue) 1 M
Nup50 (residues 1-109, K3E/R4D) or with (green) 1 M
Nup50 (residues 1-109, R38A/R45D).
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