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PDBsum entry 1a38
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Complex (signal transduction/peptide)
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PDB id
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1a38
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Contents |
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* Residue conservation analysis
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References listed in PDB file
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Key reference
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Title
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14-3-3zeta binds a phosphorylated raf peptide and an unphosphorylated peptide via its conserved amphipathic groove.
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Authors
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C.Petosa,
S.C.Masters,
L.A.Bankston,
J.Pohl,
B.Wang,
H.Fu,
R.C.Liddington.
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Ref.
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J Biol Chem, 1998,
273,
16305-16310.
[DOI no: ]
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PubMed id
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Abstract
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14-3-3 proteins bind a variety of molecules involved in signal transduction,
cell cycle regulation and apoptosis. 14-3-3 binds ligands such as Raf-1 kinase
and Bad by recognizing the phosphorylated consensus motif, RSXpSXP, but must
bind unphosphorylated ligands, such as glycoprotein Ib and Pseudomonas
aeruginosa exoenzyme S, via a different motif. Here we report the crystal
structures of the zeta isoform of 14-3-3 in complex with two peptide ligands: a
Raf-derived phosphopeptide (pS-Raf-259, LSQRQRSTpSTPNVHMV) and an
unphosphorylated peptide derived from phage display (R18, PHCVPRDLSWLDLEANMCLP)
that inhibits binding of exoenzyme S and Raf-1. The two peptides bind within a
conserved amphipathic groove on the surface of 14-3-3 at overlapping but
distinct sites. The phosphoserine of pS-Raf-259 engages a cluster of basic
residues (Lys49, Arg56, Arg60, and Arg127), whereas R18 binds via the
amphipathic sequence, WLDLE, with its two acidic groups coordinating the same
basic cluster. 14-3-3 is dimeric, and its two peptide-binding grooves are
arranged in an antiparallel fashion, 30 A apart. The ability of each groove to
bind different peptide motifs suggests how 14-3-3 can act in signal transduction
by inducing either homodimer or heterodimer formation in its target proteins.
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Figure 1.
Fig. 1. Stereodiagram of the 14-3-3  monomer
structure showing electron density for the pS-Raf-259 and R18
peptides. Helices forming the amphipathic groove are in white.
In red is an F[o]  F[c] map
contoured at 2  calculated
for crystals soaked in the a) pS-Raf-259 or b) R18 peptide. The
maps are at 3.6 (A) and 3.35 Å (B) resolution using phases
calculated from the protein model before inclusion of peptide
atoms and improved by 4-fold noncrystallographic symmetry
averaging, histogram matching, and solvent flattening in DM
(25). The figure was produced with Bobscript (30) and Raster3D
(31).
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Figure 3.
Fig. 3. Amphipathicity and sequence conservation of the
peptide-binding site. A, space-filling model of 14-3-3 with
residues defining the amphipathic groove colored by side chain
type: hydrophobic (green), polar (dark gray), acidic (red), and
basic (blue). The pS-Raf-259 peptide backbone with its
phosphoserine side chain is shown in yellow. Asp or Glu
substitutions leading to reduced Raf binding3 (19) are marked
with * (strong effect) or with ± (weak effect). B, the
concave inner surface of 14-3-3 with residues invariant across
30 eukaryotic species in red (see also Table II). The R18
peptide is shown in green. None of the residues solvent-exposed
on the rear, convex surface are invariant (not shown). C,
close-up view of residues from helices  3, 5, 7, and 9 forming
the amphipathic groove. All residues exposed in the groove are
labeled except Gly53 and Gly169. The viewing orientation and
coloring scheme are as in A. Residues boxed in solid or dashed
lines correspond to those marked in A by * or ±,
respectively. D, schematic of a 14-3-3 dimer with
helices as cylinders showing bound Raf peptides with their
phosphoserine side chains. The two peptides are oriented in an
antiparallel fashion. The view is rotated by 90 ° around a
horizontal axis compared with A-C, so that the dyad axis lies
vertically in the plane of the page. The figure was produced
with Molscript (32) and Raster3D (31).
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The above figures are
reprinted
by permission from the ASBMB:
J Biol Chem
(1998,
273,
16305-16310)
copyright 1998.
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Secondary reference #1
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Title
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14-3-3zeta binds a phosphorylated raf peptide and an unphosphorylated peptide via its conserved amphipathic groove.
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Authors
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C.Petosa,
S.C.Masters,
L.A.Bankston,
J.Pohl,
B.Wang,
H.Fu,
R.C.Liddington.
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Ref.
|
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J Biol Chem, 1998,
273,
16305-16310.
[DOI no: ]
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PubMed id
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Figure 1.
Fig. 1. Stereodiagram of the 14-3-3 monomer
structure showing electron density for the pS-Raf-259 and R18
peptides. Helices forming the amphipathic groove are in white.
In red is an F[o] F[c] map
contoured at 2 calculated
for crystals soaked in the a) pS-Raf-259 or b) R18 peptide. The
maps are at 3.6 (A) and 3.35 Å (B) resolution using phases
calculated from the protein model before inclusion of peptide
atoms and improved by 4-fold noncrystallographic symmetry
averaging, histogram matching, and solvent flattening in DM
(25). The figure was produced with Bobscript (30) and Raster3D
(31).
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Figure 3.
Fig. 3. Amphipathicity and sequence conservation of the
peptide-binding site. A, space-filling model of 14-3-3 with
residues defining the amphipathic groove colored by side chain
type: hydrophobic (green), polar (dark gray), acidic (red), and
basic (blue). The pS-Raf-259 peptide backbone with its
phosphoserine side chain is shown in yellow. Asp or Glu
substitutions leading to reduced Raf binding3 (19) are marked
with * (strong effect) or with ± (weak effect). B, the
concave inner surface of 14-3-3 with residues invariant across
30 eukaryotic species in red (see also Table II). The R18
peptide is shown in green. None of the residues solvent-exposed
on the rear, convex surface are invariant (not shown). C,
close-up view of residues from helices 3, 5, 7, and 9 forming
the amphipathic groove. All residues exposed in the groove are
labeled except Gly53 and Gly169. The viewing orientation and
coloring scheme are as in A. Residues boxed in solid or dashed
lines correspond to those marked in A by * or ±,
respectively. D, schematic of a 14-3-3 dimer with
helices as cylinders showing bound Raf peptides with their
phosphoserine side chains. The two peptides are oriented in an
antiparallel fashion. The view is rotated by 90 ° around a
horizontal axis compared with A-C, so that the dyad axis lies
vertically in the plane of the page. The figure was produced
with Molscript (32) and Raster3D (31).
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The above figures are
reproduced from the cited reference
with permission from the ASBMB
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