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PDBsum entry 1g4b
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Chaperone/hydrolase
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PDB id
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1g4b
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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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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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Abstract
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BACKGROUND: The bacterial heat shock locus HslU ATPase and HslV peptidase
together form an ATP-dependent HslVU protease. Bacterial HslVU is a homolog of
the eukaryotic 26S proteasome. Crystallographic studies of HslVU should provide
an understanding of ATP-dependent protein unfolding, translocation, and
proteolysis by this and other ATP-dependent proteases. RESULTS: We present a 3.0
A resolution crystal structure of HslVU with an HslU hexamer bound at one end of
an HslV dodecamer. The structure shows that the central pores of the ATPase and
peptidase are next to each other and aligned. The central pore of HslU consists
of a GYVG motif, which is conserved among protease-associated ATPases. The
binding of one HslU hexamer to one end of an HslV dodecamer in the 3.0 A
resolution structure opens both HslV central pores and induces asymmetric
changes in HslV. CONCLUSIONS: Analysis of nucleotide binding induced
conformational changes in the current and previous HslU structures suggests a
protein unfolding-coupled translocation mechanism. In this mechanism, unfolded
polypeptides are threaded through the aligned pores of the ATPase and peptidase
and translocated into the peptidase central chamber.
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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
reprinted
by permission from Cell Press:
Structure
(2001,
9,
177-184)
copyright 2001.
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Secondary reference #1
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Title
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Purification and characterization of the heat shock proteins hslv and hslu that form a new ATP-Dependent protease in escherichia coli.
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Authors
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S.J.Yoo,
J.H.Seol,
D.H.Shin,
M.Rohrwild,
M.S.Kang,
K.Tanaka,
A.L.Goldberg,
C.H.Chung.
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Ref.
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J Biol Chem, 1996,
271,
14035-14040.
[DOI no: ]
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PubMed id
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Figure 2.
Fig. 2. Elution of HslV from a Sephacryl S-300 column. The
proteins (18 mg) obtained from DEAE-Sepharose column were loaded
on a Sephacryl S-300 column (2 × 80 cm) equilibrated with
buffer B containing 0.1 M NaCl. Fractions of 2 ml were
collected, and aliquots of them were assayed for the hydrolysis
of Cbz-Gly-Gly-Leu-AMC (A) or electrophoresed as described in
the legend to Fig. 1 (B). The closed circles indicate the
peptide hydrolysis, and the dotted line shows the protein
profile.
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Figure 7.
Fig. 7. Determination of kinetic parameters for ATP
hydrolysis by HslU in the presence and absence of HslV. ATP
hydrolysis was assayed by incubating 1 µg of HslU and
increasing amounts of ATP in the absence (  circle )
and presence (  ) of 1
µg of HslV for 1 h at 37 °C.
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The above figures are
reproduced from the cited reference
with permission from the ASBMB
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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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