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PDBsum entry 1h6h
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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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The crystal structure of the px domain from p40(phox) bound to phosphatidylinositol 3-Phosphate.
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Authors
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J.Bravo,
D.Karathanassis,
C.M.Pacold,
M.E.Pacold,
C.D.Ellson,
K.E.Anderson,
P.J.Butler,
I.Lavenir,
O.Perisic,
P.T.Hawkins,
L.Stephens,
R.L.Williams.
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Ref.
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Mol Cell, 2001,
8,
829-839.
[DOI no: ]
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PubMed id
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Abstract
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More than 50 human proteins with a wide range of functions have a 120 residue
phosphoinositide binding module known as the PX domain. The 1.7 A X-ray crystal
structure of the PX domain from the p40(phox) subunit of NADPH oxidase bound to
PtdIns(3)P shows that the PX domain embraces the 3-phosphate on one side of a
water-filled, positively charged pocket and reveals how 3-phosphoinositide
specificity is achieved. A chronic granulomatous disease (CGD)-associated
mutation in the p47(phox) PX domain that abrogates PtdIns(3)P binding maps to a
conserved Arg that does not directly interact with the phosphoinositide but
instead appears to stabilize a critical lipid binding loop. The SH3 domain
present in the full-length protein does not affect soluble PtdIns(3)P binding to
the p40(phox) PX domain.
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Figure 5.
Figure 5. PtdIns(3)P Bound to the p40^phoxPX Domain(A)
σ[A]-weighted electron density for the di-C4-PtdIns(3)P and
some of the surrounding amino acids. The 2mF[o]-DF[c] map was
contoured at 1 σ and rendered with BOBSCRIPT (Esnouf, 1999).(B)
A view of the PtdIns(3)P binding pocket. Interacting residues
are drawn as cyan sticks, and hydrogen bonds are shown by dashed
lines. The inositol ring also makes hydrophobic contacts with
the side chain of Tyr59, and the diacylglycerol moiety makes
hydrophobic contacts with the side chain of Tyr94. In addition,
the Arg57/Asp139 salt link is shown
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Figure 7.
Figure 7. Comparison of Three Phosphoinositide Binding
ModulesThe phosphoinositide binding sites in the EEA1 FYVE
domain (PDB: 1hyi) (Kutateladze and Overduin, 2001), the
p40^phox PX domain, and the Grp1 PH domain (PDB: 1fgy) (Lietzke
et al., 2000) are shown. To illustrate the binding
environments, the structures were superimposed using their
inositol rings. The molecular surfaces are colored by
electrostatic potential.(A) The EEA1 FYVE domain with PtdIns(3)P
bound in a basic pocket.(B) The p40^phox PX domain. The
PtdIns(3)P sits in a basic pocket that interacts with one face
of the ring, while the other face is exposed to the solvent.(C)
The Grp1 PH domain bound to Ins(1,3,4,5)P[4]. As with the
p40^phox PX domain, only one face of the inositol ring interacts
with a basic patch on the surface of the domain
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The above figures are
reprinted
by permission from Cell Press:
Mol Cell
(2001,
8,
829-839)
copyright 2001.
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Secondary reference #1
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Title
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Solution structure of the px domain, A target of the sh3 domain.
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Authors
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H.Hiroaki,
T.Ago,
T.Ito,
H.Sumimoto,
D.Kohda.
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Ref.
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Nat Struct Biol, 2001,
8,
526-530.
[DOI no: ]
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PubMed id
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Figure 2.
Figure 2. In vitro interactions of the p47 PX and the p47
C-terminal SH3. a, p47 SH3 domains (the N-terminal SH3 and
the C-terminal and its mutant SH3) fused to GST were tested for
interactions with a His[6]-tagged p47 PX. b, p47 C-terminal SH3
(p47 SH3(C)) fused to GST was tested for interaction with
mutants of the His[6]-tagged p47 PX. The double-headed arrow
indicates the position of the His[6]-tagged p47 PX, which was
copurified with the GST-SH3s attached to glutathione beads. Only
the interaction between the wild type p47 PX and the wild type
p47 C-terminal SH3 was detected. Figure shown is representative
of at least three experiments.
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Figure 3.
Figure 3. Structure of the p47 PX domain. a, Overlay of the
20 final NMR structures. The residues used for superimposing the
different structures are colored in gray and the other residues
in cyan. b, Ribbon representation of the best minimized NMR
structure. The sheet and helix elements are labeled and colored
green and blue, respectively. The proline-rich motif, residues
70 -76, is colored yellow.
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The above figures are
reproduced from the cited reference
with permission from Macmillan Publishers Ltd
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