|
Figure 1.
Fig. 1. Structure of the Miz-1 POZ domain. (a) Ribbon
representation of the Miz-1 POZ domain tetramer. Subunits A–D
are indicated in different colours. The secondary structure
elements of the A-chain and of the A:D interface are shown. The
structure of the BCL6 POZ domain (PDB accession code 1r28) is
shown for comparison; the box indicates the β1–β5′
beta-sheet of the BCL6 dimerisation interface that is absent in
Miz-1 (positions on the partner chain of BCL6 are indicated with
primes). The Miz-1 POZ domain (residues 2–115) was expressed
as a glutathione S-transferase fusion protein in E. coli
BL21(DE3)pLysS. The glutathione S-transferase tag was removed by
cleavage with PreScission protease, and the Miz-1 POZ domain
fragment was purified by chromatography on Q-Sepharose, Resource
Q and Supadex 75. Crystals of the purified Miz-1 POZ domain were
obtained by hanging-drop vapour diffusion against 20%
polyethylene glycol 3350; 20% polyethylene glycol 400; 200 mM
MgCl[2]; and 100 mM Hepes (pH 7.5). Details of plasmid
construction, protein purification, X-ray data collection and
structure determination are reported in Supplementary
Data.^[20.]^, ^[21.]^, ^[22.]^, ^[23.]^, ^[24.]^, ^[25.]^,
^[26.]^ and ^[27.] (b) Superposition of POZ domain Cα atoms for
Miz-1, BCL6, PLZF and LRF. Stereoimage of the superpositions of
POZ domain dimers (BCL6, accession code 1r28; PLZF, accession
code 1buo; LRF, accession code 2nn2; the Miz-1 dimer corresponds
to chains A and B). Miz-1 (green), BCL6 (cyan), LRF (magenta),
PLZF (yellow).
|
