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Figure 3.
Figure 3. Accessible Hydrophobic Residues in the Predicted
Coiled Coil Are Critical for Rab Binding
(A) Helical wheel
projection of a coiled coil predicted for GCC185 residues
1579–1606. Residues in registers “a–g” were predicted by
the Paircoil program. Residues at positions “a” and “d”
lie in the dimer interface. Boxed residues are candidates for
binding interactions with Rab GTPases.
(B and C) Effect of
alanine substitutions on Rab binding. Reactions contained
wild-type or mutant GST-C-110 ([B] 3 μM, [C] 2 μM) and
^35S-GTPγS-preloaded GTPases ([B] 170 pmol Rab9-His, [C] 190
pmol His-Rab6). Data are mean ± SD.
(D) Mass
determination of untagged RBD-87 I1588A/L1595A by multiple angle
static light scattering. The gel filtration elution profile of
the protein (black line) and molecular mass (gray line) are
shown. Polydispersity of the peak was 1.001.
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