 |
PDBsum entry 2rm0
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Transcription
|
PDB id
|
|
|
|
2rm0
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
References listed in PDB file
|
|
|
Key reference
|
|
|
Title
|
|
Structural characterization of a new binding motif and a novel binding mode in group 2 ww domains.
|
|
|
Authors
|
|
X.Ramirez-Espain,
L.Ruiz,
P.Martin-Malpartida,
H.Oschkinat,
M.J.Macias.
|
|
|
Ref.
|
|
J Mol Biol, 2007,
373,
1255-1268.
[DOI no: ]
|
|
|
PubMed id
|
|
|
|
|
|
Abstract
|
|
|
Formin homology 1 (FH1), is a long proline-rich region of formins, shown to bind
to five WW containing proteins named formin binding proteins (FBPs). FH1 has
several potential binding regions but only the PPLPx motif and its interaction
with FBP11WW1 has been characterized structurally. To detect whether additional
motifs exist in FH1, we synthesized five peptides and investigated their
interaction with FBP28WW2, FBP11WW1 and FBP11WW2 domains. Peptides of sequence
PTPPPLPP (positive control), PPPLIPPPP and PPLIPPPP (new motifs) interact with
the domains with micromolar affinity. We observed that FBP28WW2 and FBP11WW2
behave differently from FBP11WW1 in terms of motif selection and affinity, since
they prefer a doubly interrupted proline stretch of sequence PPLIPP. We
determined the NMR structure of three complexes involving the FBP28WW2 domain
and the three ligands. Depending on the peptide under study, the domain
interacts with two proline residues accommodated in either the XP or the XP2
groove. This difference represents a one-turn displacement of the domain along
the ligand sequence. To understand what drives this behavior, we performed
further structural studies with the FBP11WW1 and a mutant of FBP28WW2 mimicking
the XP2 groove of FBP11WW1. Our observations suggest that the nature of the XP2
groove and the balance of flexibility/rigidity around loop 1 of the domain
contribute to the selection of the final ligand positioning in fully independent
domains. Additionally, we analyzed the binding of a double WW domain region,
FBP11WW1-2, to a long stretch of FH1 using fluorescence spectroscopy and NMR
titrations. With this we show that the presence of two consecutive WW domains
may also influence the selection of the binding mode, particularly if both
domains can interact with consecutive motifs in the ligand. Our results
represent the first observation of protein-ligand recognition where a pair of WW
and two consecutive motifs in a ligand participate simultaneously.
|
|
|
|
|
|
|
|
Figure 4.
Figure 4. Solution structure of FBP28WW in complex with the
PPPLIPPPP peptide. (a) Stereo view of the best-fit backbone (N,
C^α, C′) superposition of the ten lowest energy structures
after water refinement with the ligand (domain in blue and
ligand in red). (b) Left: Surface representation of the lowest
energy structure (ligand in brown). Right: Cartoon
representation with the secondary structural elements of the
domain shown in green, residues participating in contacts in
dark yellow and ligand in brown.
|
|
Figure 5.
Figure 5. Solution structure of FBP28WW in complex with the
PPLIPPPP peptide. (a) Stereo view of the best-fit backbone (N,
C^α, C′) superposition of the ten lowest energy structures
after water refinement with the ligand (domain in blue and
ligand in red). (b) Left: Corresponding surface representation
of the lowest energy structure (ligand in dark green). Right:
Cartoon representation with the secondary structural elements of
the domain shown in green, residues participating in contacts in
dark yellow and ligand in dark green.
|
|
|
|
|
|
The above figures are
reprinted
by permission from Elsevier:
J Mol Biol
(2007,
373,
1255-1268)
copyright 2007.
|
|
|
|
|
|
|
