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Figure 3.
FIGURE 3. Purification and crystallization of the PPAR  LBD
complexed with a PGC-1  fragment. A, a
schematic representation showing the PGC-1 protein and its two
receptor-interacting motifs (ID1 and ID2). The PGC-1 fragment
(101-220) that includes both ID1 and ID2 motifs was used in
cocrystallization with PPAR . B, purification of
the PPAR LBD and PGC-1 complex.
The proteins of PPAR and PGC-1 were
purified separately and complexed using excess PGC-1 in the
presence of rosiglitazone. The PPAR ·PGC-1 complex
and PGC-1 alone were separated by
gel filtration. The complex was eluted in the first peak and
collected and concentrated to 10 mg/ml for the crystallization
trial. C, the protein complex samples shown on an SDS gel. The
molecular mass markers are shown in the KD lane (kilodaltons).
Lanes 1-9 are fractions from the gel filtration column
corresponding to the two peaks in B, from left to right. D,
crystals of the PPAR ·rosiglitazone·PGC-1
complex.
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