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PDBsum entry 1im2
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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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Structure of haemophilus influenzae hslu protein in crystals with one-Dimensional disorder twinning.
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Authors
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C.B.Trame,
D.B.Mckay.
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Ref.
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Acta Crystallogr D Biol Crystallogr, 2001,
57,
1079-1090.
[DOI no: ]
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PubMed id
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Abstract
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The structure of the Haemophilus influenzae HslU protein, a molecular chaperone
of the Clp/Hsp100 family, has been solved to 2.3 A by molecular replacement
using a model of the homologous Escherichia coli protein. The crystals in which
the structure was solved have an unusual twinning, or one-dimensional disorder,
in which each successive crystal-packing layer is displaced laterally relative
to the one below it. A model for the twinning and an algorithm for detwinning
the data are described. It is known from other work that when the HslU hexamer
binds its cognate protease HslV, the carboxy-terminal helices of HslU protomers
distend and bind between HslV subunits. Comparison of HslU alone with its
structure in the HslUV complex reveals several conserved amino-acid residues
whose side-chain interactions differ between the two structures, suggesting that
they may be part of a conformational switch that facilitates the release of the
HslU carboxy-terminal helices when HslV binds.
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Figure 5.
Figure 5 Representative electron-density map around the
nucleotide-binding site. Stereoview of a simulated-annealing
omit map (green, contoured at 4.2  )
in which ADP and Arg394 were omitted from the model used in
refinement. Protein is shown as a ribbon diagram; ADP and
residues Arg394 and Lys63 are shown in ball-and-stick
representation. Distances from the non-bonded guanidinium N
atoms of Arg394 to the nearest phosphate O atom are 2.8 and 2.5
Å; distance from the amino group of Lys63 to the nearest
phosphate O atom is 2.8 Å.
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Figure 9.
Figure 9 Electrostatic surface potentials of HslV and HslU.
Positions of some amino-acid residues are labeled in each
figure. (a) View looking down the sixfold axis of the HslV
dodecamer from the HslUV complex (Sousa et al., 2000[Sousa, M.
C., Trame, C. B., Tsuruta, H., Wilbanks, S. M., Reddy, V. S. &
McKay, D. B. (2000). Cell, 103, 633-643.]). Arrows indicate the
electropositive grooves into which the carboxy-terminal helices
of HslU intercalate. (b) Carboxy-terminal helix of one protomer
of HslU in its conformation from the HslUV complex. (c) Side
view showing the interface between the carboxy-terminal helix of
HslU, shown as a ball-and-stick model, and the groove between
two HslV protomers. Electrostatic potentials were computed with
the program GRASP (Nicholls & Honig, 1991[Nicholls, A. & Honig,
B. J. (1991). J. Comput. Chem. 12, 435-445.]), using a
dielectric constant of 2.0 for the interior of the protein and
80.0 for the solvent area and an effective ionic strength
equivalent to 1.0 M salt.
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The above figures are
reprinted
by permission from the IUCr:
Acta Crystallogr D Biol Crystallogr
(2001,
57,
1079-1090)
copyright 2001.
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Secondary reference #1
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Title
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The structures of hsiu and the ATP-Dependent protease hsiu-Hsiv.
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Authors
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M.Bochtler,
C.Hartmann,
H.K.Song,
G.P.Bourenkov,
H.D.Bartunik,
R.Huber.
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Ref.
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Nature, 2000,
403,
800-805.
[DOI no: ]
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PubMed id
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Figure 1.
Figure 1: Summary of the three crystal forms (a-c) that were
used for structure determination. Subunits in the respective
asymmetric units are numbered 1-6.
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Figure 2.
Figure 2: Comparison of HsIU and NSF main chains. a,
Superposition of the ligand-bound (coloured) and free (white)
HslU forms. Chains 1 and 2 of the P321 crystals (see Fig. 1c)
are shown. The N domains (shown in green and red) have been
superimposed (r.m.s.d. C  bond
lengths = 0.5 Å for the central  -sheet,
r.m.s.d. C bond
lengths = 1.2 Å for the whole domain). For clarity, the N and I
domains of the free form have been omitted. b, Stereo diagram of
NSF D2.
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The above figures are
reproduced from the cited reference
with permission from Macmillan Publishers Ltd
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Secondary reference #2
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Title
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Mutational studies on hslu and its docking mode with hslv.
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Authors
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H.K.Song,
C.Hartmann,
R.Ramachandran,
M.Bochtler,
R.Behrendt,
L.Moroder,
R.Huber.
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Ref.
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Proc Natl Acad Sci U S A, 2000,
97,
14103-14108.
[DOI no: ]
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PubMed id
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Figure 2.
Fig. 2. Representation of the electrostatic potential
surfaces of HslV (Left) and HslU (Center) involved in the EM
mode of docking. Negatively charged regions are in red, and
positively charged regions are in blue. Sites of mutations in
the HslU (Right). Numbers 1 (green) and 3 (pink) mark sites of
pentaglycine insertions after residues 264 and 387 as well as
changes of surface charges (E266Q; E266Q/E385K), 2 (blue) marks
the site of introduction of a bulky side chain (I312W), and 4
(red) marks the site of a charge reversal (E436K/D437K). The
hexamer pore is colored in yellow. This figure was drawn by
using GRASP (28).
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Figure 3.
Fig. 3. Sites of mutations in the hexamer pore.
Side-chain atoms (yellow) are shown only in one subunit for
clarity. Mutation sites in the hexamer pore are colored in pink.
Top view of HslU (Left). Side view of the central pore of HslU
hexamer (Right). Two subunits from the ring nearest to the
reader are removed to expose the interior. This figure was drawn
by using GRASP (28).
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Secondary reference #3
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Title
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Crystal and solution structures of an hsluv protease-Chaperone complex.
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Authors
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M.C.Sousa,
C.B.Trame,
H.Tsuruta,
S.M.Wilbanks,
V.S.Reddy,
D.B.Mckay.
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Ref.
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Cell, 2000,
103,
633-643.
[DOI no: ]
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PubMed id
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Figure 1.
Figure 1. Representative Electron Density MapsStereo views
of F[o] − F[c] simulated annealing omit maps, computed with
phases calculated from models in which the atoms of interest
were deleted from the model used in refinement.(A) the ATP
binding site of HslU, contoured at 5σ. Protein is shown as a
ribbon diagram; ATP from the final HslUV model (average B factor
29.3) is shown as a ball and stick representation.(B)
Carboxy-terminal segment of HslU (average B factor 119.1),
contoured at 3σ (magenta) and 6σ (cyan). Residues of HslU
which were omitted are shown in green, oriented with the
carboxy-terminal Leu-444 at the bottom of the figure;
neighboring residues of HslV are shown in standard colors
(oxygen, red; nitrogen, blue; carbon, gray). Figure was prepared
with BOBSCRIPT ([7 and 8]). The rendering and stereo pair
generation of all figures was done with RASTER3D ( [25]) and
IMAGEMAGIK
(http://www.wizards.dupont.com/cristy/ImageMagick.html).
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Figure 6.
Figure 6. Conformational Changes around the Catalytic Site
of HslVStereo ribbon drawing of the active site region. The
HslUV structure is colored green. The segment of uncomplexed
HslV that differs substantially from the complex (see Figure 3A)
is colored magenta. Selected residue side chains and polypeptide
backbone are shown in the ball and stick representation.
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The above figures are
reproduced from the cited reference
with permission from Cell Press
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