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PDBsum entry 1hyi
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Endocytosis/exocytosis
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PDB id
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1hyi
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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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Structural mechanism of endosome docking by the fyve domain.
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Authors
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T.Kutateladze,
M.Overduin.
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Ref.
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Science, 2001,
291,
1793-1796.
[DOI no: ]
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PubMed id
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Abstract
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The recruitment of trafficking and signaling proteins to membranes containing
is mediated by FYVE domains. Here,
the solution structure of the FYVE domain of the early endosome antigen 1
protein (EEA1) in the free state was compared with the structures of the domain
complexed with PtdIns(3)P and mixed micelles. The multistep binding mechanism
involved nonspecific insertion of a hydrophobic loop into the lipid bilayer,
positioning and activating the binding pocket. Ligation of PtdIns(3)P then
induced a global structural change, drawing the protein termini over the bound
phosphoinositide by extension of a hinge. Specific recognition of the
3-phosphate was determined indirectly and directly by two clusters of conserved
arginines.
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Figure 1.
Fig. 1. Solution structures of the EEA1 FYVE domain. (A) The C
 traces of
20 structures are shown that comprise EEA1 residues 1346 to 1410
in the lipid-free form, with the  strands,
helix, hinge, membrane insertion loop, and zinc ions colored
green, magenta, cyan, brown, and orange, respectively. (B)
Ribbon diagram of the structure closest to the average. The 1, 2, 3, and 4 strands
and 1 helix
consist of residues 1370 to 1372, 1379 to 1381, 1387 to 1391,
1396 to 1400, and 1402 to 1408, respectively. The two zinc ions
(Zn1 and Zn2) are coordinated by eight cysteines whose side
chains are shown in yellow. (C) The C traces of
20 PtdIns(3)P-bound structures are depicted as in (A). The
PtdIns(3)P head group of the average structure is shown, with C
and P atoms in yellow and O atoms in red. (D) Superposition of
the PtdIns(3)P-bound and free structures, which are colored and
gray, respectively. The view has been rotated slightly to reveal
the structural differences.
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Figure 3.
Fig. 3. Phosphate group recognition by arginines. (A) The H
 resonances
of Arg1369 and Arg1374 shift downfield upon PtdIns(3)P
interaction (lower panel), indicating the formation of new
hydrogen bonds. The H[2]N  resonances
of the Arg1370 and Arg1374 become resolved only in the presence
of PtdIns(3)P (upper panel), indicating that the guanidino group
rotation is impeded by PtdIns(3)P coordination. Two regions of
five 1H-15N correlation spectra of the FYVE domain (1 mM) with
PtdIns(3)P concentrations of 0, 0.1, 0.5, 1, and 5 mM are
superimposed. Arginine's structure is shown in the upper right.
(B) PtdIns(3)P moieties recognized by the FYVE domain are
indicated by absolute changes in the 1H, 13C, and 31P chemical
shifts of PtdIns(3)P induced by addition of equimolar FYVE
domain. Atoms are numbered according to the positions indicated
in (C). (C) Model of dibutanoyl PtdIns(3)P, with atom colors
based on chemical shift perturbations induced by addition of
equimolar FYVE domain, as indicated in (B).
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The above figures are
reprinted
by permission from the AAAs:
Science
(2001,
291,
1793-1796)
copyright 2001.
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