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(+ 2 more)
174 a.a.
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408 a.a.
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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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Nucleotide-Dependent conformational changes in a protease-Associated atpase hsiu.
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Authors
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J.Wang,
J.J.Song,
I.S.Seong,
M.C.Franklin,
S.Kamtekar,
S.H.Eom,
C.H.Chung.
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Ref.
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Structure, 2001,
9,
1107-1116.
[DOI no: ]
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PubMed id
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Abstract
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BACKGROUND: The bacterial heat shock locus ATPase HslU is an AAA(+) protein that
has structures known in many nucleotide-free and -bound states. Nucleotide is
required for the formation of the biologically active HslU hexameric assembly.
The hexameric HslU ATPase binds the dodecameric HslV peptidase and forms an
ATP-dependent HslVU protease. RESULTS: We have characterized four distinct HslU
conformational states, going sequentially from open to closed: the empty, SO(4),
ATP, and ADP states. The nucleotide binds at a cleft formed by an alpha/beta
domain and an alpha-helical domain in HslU. The four HslU states differ by a
rotation of the alpha-helical domain. This classification leads to a correction
of nucleotide identity in one structure and reveals the ATP hydrolysis-dependent
structural changes in the HslVU complex, including a ring rotation and a
conformational change of the HslU C terminus. This leads to an amended protein
unfolding-coupled translocation mechanism. CONCLUSIONS: The observed
nucleotide-dependent conformational changes in HslU and their governing
principles provide a framework for the mechanistic understanding of other AAA(+)
proteins.
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Figure 5.
Figure 5. The Binding of ATP-Bound HslU Opens the Central
Pore of HslV(a) Uncomplexed HslV has a closed pore [27].(b) The
binding of the ATP-bound HslU opens the central pore of HslV
with an average diameter of 19.3Å [27]. The HslU C terminus is
shown in magenta. The insertion of the C terminus and relative
twisting ring rotation may be responsible for the pore opening
in a "twist-and-open" mechanism. Two conserved arginines, Arg-86
and Arg-89, are also shown. There are two more positively
charged residues, which are not shown, nearby at the pore of
HslV (Arg-83 and Lys-90)
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The above figure is
reprinted
by permission from Cell Press:
Structure
(2001,
9,
1107-1116)
copyright 2001.
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Secondary reference #1
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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 #2
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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 #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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Secondary reference #4
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Title
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Crystal structures of the hslvu peptidase-Atpase complex reveal an ATP-Dependent proteolysis mechanism.
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Authors
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J.Wang,
J.J.Song,
M.C.Franklin,
S.Kamtekar,
Y.J.Im,
S.H.Rho,
I.S.Seong,
C.S.Lee,
C.H.Chung,
S.H.Eom.
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Ref.
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Structure, 2001,
9,
177-184.
[DOI no: ]
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PubMed id
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Figure 1.
Figure 1. The Structures of HslVU(a) A composite-omit
electron density map (cyan, contoured at 1s) at 3.0 Å resolution
reveals that the bound dADP (yellow) is in an anti conformation,
not syn, as in a previously determined structure (AMPPNP,
magenta). This map was generated before dADP was built into the
model.(b) The HslVU complex in the asymmetric U[6]V[6]V[6]
configuration. Parts of HslU domain I could not be built into
the final electron density and are indicated by spheres for
their approximate locations.(c) The HslVU structure in the
symmetric U[6]V[6]V[6]U[6] configuration. The orientation of the
complexes in (1b) and (1c) differs by 30°
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The above figure is
reproduced from the cited reference
with permission from Cell Press
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Secondary reference #5
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Title
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Docking of components in a bacterial complex.
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Authors
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T.Ishikawa,
M.R.Maurizi,
D.Belnap,
A.C.Steven.
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Ref.
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Nature, 2000,
408,
667-668.
[DOI no: ]
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PubMed id
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Figure 1.
Figure 1: Averaged side-view projections of HslVU complexes.
a-c, Electron micrographs of complexes formed in 50 mM Tris-
HCl, pH 7.5, 0.2 M KCl, 10 mM MgCl[2] and 1 mM ATP, representing
appropriate conditions for proteolytic activity. a, Negatively
stained molecules (ATP-  S
state; number of particles, N = 65; resolution, 32 Å). Note that
the proximal ring of HslU (arrowhead) is wider and more dense
than the outer ring (arrow); b, frozen-hydrated molecules (ATP-
S
state; N=250; resolution, 33 Å); c, frozen-hydrated molecules in
the AMP-PNP state, where AMP-PNP is an inactive ATP analogue (N
= 400; resolution, 33 Å). d-g, Side-view projections
calculated^6 from the crystal structure^2 but limited to 30 Å
resolution. Projections corresponding to different rotational
settings of the complex around the axis were averaged to give a
cylindrically averaged side view, as in the electron micrographs
(EMs). In d and e , HslU is in the opposite orientation from the
one in the crystal structure, whereas in f and g this
corresponds to the published orientation2. Projections shown in
e and g were created by applying a phase-contrast transfer
function (CTF; corresponding to 2.0  m
underfocus) to images in d and f , and so are more comparable to
the cryo-EMs. With or without CTF correction, it is evident that
the wider, denser ring, corresponding to the ATPase domains of
HslU, is adjacent to HslV. Arrows in e and c mark the axial
density that is missing in e but present in b and c, which we
attribute to residues 175 to 209. In e and g, I denotes the
I-domain ring, and A denotes the ATPase-domain ring. Scale bar,
100 Å.
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The above figure is
reproduced from the cited reference
with permission from Macmillan Publishers Ltd
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Secondary reference #6
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Title
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A corrected quaternary arrangement of the peptidase hslv and atpase hslu in a cocrystal structure.
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Author
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J.Wang.
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Ref.
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J Struct Biol, 2001,
134,
15-24.
[DOI no: ]
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PubMed id
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