Chen et al. 10.1073/pnas.0704525104. |
Fig. 5. (A) The omit Fo - Fc difference map at 2.5 s. The structure of c-JMJD2A was used as initial model for Fc (1), a 2.0-Ã… experimental reflection data set was used as Fo, and the map was calculated with the Crystallography and NMR System (2). There is one break density at residue His-39. The side chains of residues Lys-37, His-39, and Arg-42 are disordered. The trimethyl group is resolved and very well defined in the map as well as the side chains of Thr-32, Val-35, Arg-40, and Trp-41. (B) The omit map of the monomethyl peptide.
1. Chen Z, Zang J, Whetstine J, Hong X, Davrazou F, Kutateladze TG, Simpson M, Mao Q, Pan CH, Dai S, et al. (2006) Cell 125:691-702.
2. Brunger AT, Adams PD, Clore GM, DeLano WL, Gros P, Grosse-Kunstleve RW, Jiang JS, Kuszewski J, Nilges M, Pannu NS, et al. (1998) Acta Crystallogr D 54:905-921.
Fig. 6. The conformation change upon peptide binding is small. The most significant change occurs at the C-terminal coil region between a9 and a10. Upon the binding of the peptide, the coil region is bent toward the catalytic center, allowing one additional hydrogen bond to form between the amide group of Val-35 and the carboxyl group of Asp-311 (Fig. 2A).
Fig. 7. The overall structure of two complexes in one asymmetric unit shows one complex with 14 peptide residues (molecule B, blue) and the other complex with 17 peptide residues (molecule A, yellow). The C-terminal regions of both peptides crash into each other at the boundary between the complexes.
Fig. 8. A comparison between the complex of FIH with a CAD peptide [residues 795-806; Protein Data Bank ID code 1H2K (1)] and that of c-JMJD2A with the H3K36me3 peptide (residues 27-43). (A) The complex structure of c-JMJD2A and the H3K36me3 peptide. (B) The complex structure of FIH and a CAD peptide. (C) Superposition of the two complexes.
1. Elkins JM, Hewitson KS, McNeill LA, Seibel JF, Schlemminger I, Pugh CW, Ratcliffe PJ, Schofield CJ (2003) J Biol Chem 278:1802-1806.
Fig. 9. Critical residues involved in the interactions with the peptides. A structure-based sequence alignment of members of the JMJD2 families (JMJD2A, B, C, and D). The potential peptide-binding residues are colored red, and the residues involved in methyl-group binding are colored blue. a, b, and 3.10H denote a-helices, b-strands, and 3.10 helices, respectively.
Fig. 10. The binding properties of different mutant variants of c-JMJD2A and c-JMJD2D to peptides measured in BIAcore experiments. Two of the mutants (double mutant G133A/G138A and double mutant G165A/G170A) do not bind to the peptide, whereas three of the mutants (D135A, Y177F, and K241R) bound to the trimethylated peptide but did not show any demethylation activity.