 | Acta Crystallographica Section D Acta Crystallographica Section D BIOLOGICAL CRYSTALLOGRAPHY |
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![[Figure 1]](mh5164fig1mag.jpg)
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Figure 1
In vivo and in vitro interaction between Stardust/Pals1 and Crumbs/Crb1. (a) Schematic for Pals1 and Crumbs domain organization. EGF, epidermal growth factor; LamAG, laminin AG-like; TM, transmembrane; ICD, intracellular; L27, lin-2, lin-7 homology; PDZ, postsynaptic density 95, disc-large, zona occludens homology; SH3, sarcoma homology 3; GUK, guanylate kinase. The PDZ domain of Pals1 interacts with Crumbs through its ICD. (b) The amino-acid sequence of the Crb17 peptide used for biochemical experiments and for crystallization is underlined; the PDZ-binding motif is coloured. (c) D. melanogaster Crumbs (shown in red) localizes to the apical membrane of the follicle cells of egg chambers. Wild-type follicle cells (WT) are marked by the presence of green fluorescent protein (GFP). (d) Deletion of the four C-terminal residues of Crumbs (CrbΔERLI) causes a failure of Crb to localize to the apical plasma membrane of the follicle cells and disrupts polarity. Mutant cells are marked by the absence of GFP; some GFP-positive cells can be seen. (e) Representative binding curves for fluorescein-labelled Crb17 for Pals1PDZ and (f) Crb17 mutants measured by fluorescence polarization (Crb17, green; Crb17 L1405A, orange; Crb17 R1404A, pink; Crb17 E1403A, blue; Crb17 M1402A, red). The affinity between Crb17 I1406A and Pals1PDZ could not be reliably determined. (g) Summary of dissociation constants (in µM) between Crb17 and Pals1PDZ [colour-coded as in (f); Crb17 II406A, brown, Crb6, light blue] measured by Fp with fluorescein-labelled peptides. All values shown were measured in parallel from the same Pals1 preparation (except for the Crb6 peptide, which was measured using a different Pals1PDZ preparation; the Kd between Crb17 and Pals1PDZ from this preparation was 12.4 ± 2.4 µM). These data were representative of three independent experiments. |
![[Figure 1]](mh5164fig1.jpg)
| BIOLOGICAL CRYSTALLOGRAPHY |
ISSN: 1399-0047
