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(+ 6 more)
321 a.a.
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(+ 6 more)
173 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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Crystal structure of hsluv complexed with a vinyl sulfone inhibitor: corroboration of a proposed mechanism of allosteric activation of hslv by hslu.
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Authors
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M.C.Sousa,
B.M.Kessler,
H.S.Overkleeft,
D.B.Mckay.
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Ref.
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J Mol Biol, 2002,
318,
779-785.
[DOI no: ]
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PubMed id
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Abstract
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On the basis of the structure of a HslUV complex, a mechanism of allosteric
activation of the HslV protease, wherein binding of the HslU chaperone
propagates a conformational change to the active site cleft of the protease, has
been proposed. Here, the 3.1 A X-ray crystallographic structure of Haemophilus
influenzae HslUV complexed with a vinyl sulfone inhibitor is described. The
inhibitor, which reacts to form a covalent linkage to Thr1 of HslV, binds in an
"antiparallel beta" manner, with hydrogen-bond interactions between
the peptide backbone of the protease and that of the inhibitor, and with two
leucinyl side chains of the inhibitor binding in the S1 and S3 specificity
pockets of the protease. Comparison of the structure of the HslUV-inhibitor
complex with that of HslV without inhibitor and in the absence of HslU reveals
that backbone interactions would correctly position a substrate for cleavage in
the HslUV complex, but not in the HslV protease alone, corroborating the
proposed mechanism of allosteric activation. This activation mechanism differs
from that of the eukaryotic proteasome, for which binding of activators opens a
gated channel that controls access of substrates to the protease, but does not
perturb the active site environment.
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Figure 1.
Figure 1. (a) Stereo view of F[o] -F[c] electron density
map showing inhibitor bound to one subunit of HslV. Phases were
computed from HslUV model that resulted from one cycle of
simulated annealing after all inhibitor atoms were omitted from
the model. Contour levels, 5s (magenta), 12.5s (cyan). The
Figure was prepared with BOBSCRIPT[25.] and RASTER3D. [26.] (b)
Proposed structure of NLVS-HslV covalent complex with Thr1 of
HslV. [14.] Inhibitor atoms and bonds are drawn bold; HslV
atoms/bonds are drawn fine. The S1 and S3 HslV binding pockets
are shown schematically above the moieties they bind.
Orientation of inhibitor is similar to orientation in part (a).
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Figure 2.
Figure 2. Stereo views of the HslUV-NLVS inhibitor complex.
(a) View showing the hydrogen bonding interactions of the
inhibitor with HslV polypeptide backbone in the HslUV-NLVS
complex. Carbon atoms of HslV are green; carbon atoms of
inhibitor are gray; nitrogen atoms, blue; oxygen atoms, red;
sulfur atom, yellow. (b) Superposition of the substrate binding
clefts of HslUV-NLVS and the yeast proteasome with epoxomycin
(subunit K of PDB 1G65).[15.] Hydrogen bonds between protein and
inhibitor are the same as shown in part (a). Selected residues
are labeled with format "HslV#/proteasome#". Color scheme: HslV
of HslUV-NLVS complex, green; NLVS inhibitor atoms, cyan;
proteasome, red; epoxomycin inhibitor atoms, gold. (c) View
showing the inhibitor (semi-transparent CPK model) and the
binding pockets of HslUV. HslV protomer to which NLVS is
covalently attached is green; adjacent HslV protomer, yellow;
HslU, magenta; inhibitor, gray. Selected side chains are
included. Carbon atoms are the same color as corresponding
protomer; oxygen atoms, red; nitrogen atoms, blue; sulfur atoms,
cyan. (d) View showing the displacement of upper strand of
substrate binding cleft of uncomplexed HslV [9.] relative to
HslUV-NLVS; when lower segments of polypeptides are
superimposed, upper segment of uncomplexed HslV is displaced
 vert,
similar 3-4 Å from its position in the HslUV-NLVS complex.
For clarity, only selected peptide backbone and C^a atoms of the
proteins and "backbone" atoms of the inhibitor are included.
Color scheme: HslV of HslUV-NLVS complex, green; NLVS inhibitor
atoms, cyan; uncomplexed HslV, magenta. Superpositions were
computed with the program LSQMAN.[27.] The Figure was prepared
with MOLSCRIPT [28.] and RASTER3D. [26.]
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The above figures are
reprinted
by permission from Elsevier:
J Mol Biol
(2002,
318,
779-785)
copyright 2002.
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Secondary reference #1
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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 #2
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Title
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Structure of haemophilus influenzae hslv protein at 1.9 a resolution, Revealing a cation-Binding site near the catalytic site.
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Authors
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M.C.Sousa,
D.B.Mckay.
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Ref.
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Acta Crystallogr D Biol Crystallogr, 2001,
57,
1950-1954.
[DOI no: ]
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PubMed id
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Figure 1.
Figure 1 Quasi-equivalent subunit interactions within HslV. (a)
Ribbon drawing of one hexamer, looking down the pseudo-sixfold
axis. Subunits related by crystallographic twofold rotation are
shown in identical colors and denoted with a prime. Regions on
the apical helices which reveal differences in subunit-subunit
interactions are highlighted: magenta on cyan for subunit A; red
on yellow for subunit B; green on gold for subunit C. (b)
Interactions between subunits C and A, using same color coding
as in (a). (c) Interactions between subunits B and C. Figs.
1-and 2-(b) were produced with MOLSCRIPT (Kraulis, 1991[Kraulis,
P. (1991). J. Appl. Cryst. 24, 946-950.]) and Fig. 2-(a) was
produced with BOBSCRIPT (Esnouf, 1997[Esnouf, R. M. (1997). J.
Mol. Graph. 15, 132-134.], 1999[Esnouf, R. M. (1999). Acta
Cryst. D55, 938-940.]); all figures were rendered with Raster3D
(Merritt & Bacon, 1997[Merritt, E. A. & Bacon, D. J. (1997).
Methods Enzymol. 277, 505-524.]).
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The above figure is
reproduced from the cited reference
with permission from the IUCr
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Secondary reference #3
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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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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
reproduced from the cited reference
with permission from the IUCr
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