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PDBsum entry 1txe
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Transport protein
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PDB id
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1txe
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References listed in PDB file
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Key reference
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Title
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Solution structure of the active-Centre mutant i14a of the histidine-Containing phosphocarrier protein from staphylococcus carnosus.
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Authors
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A.Möglich,
B.Koch,
W.Gronwald,
W.Hengstenberg,
E.Brunner,
H.R.Kalbitzer.
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Ref.
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Eur J Biochem, 2004,
271,
4815-4824.
[DOI no: ]
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PubMed id
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Abstract
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High-pressure NMR experiments performed on the histidine-containing
phosphocarrier protein (HPr) from Staphylococcus carnosus have shown that
residue Ile14, which is located in the active-centre loop, exhibits a peculiarly
small pressure response. In contrast, the rest of the loop shows strong pressure
effects as is expected for typical protein interaction sites. To elucidate the
structural role of this residue, the mutant protein HPr(I14A), in which Ile14 is
replaced by Ala, was produced and studied by solution NMR spectroscopy. On the
basis of 1406 structural restraints including 20 directly detected hydrogen
bonds, 49 1H(N)-15N, and 25 1H(N)-1Halpha residual dipolar couplings, a well
resolved three-dimensional structure could be determined. The overall fold of
the protein is not influenced by the mutation but characteristic conformational
changes are introduced into the active-centre loop. They lead to a displacement
of the ring system of His15 and a distortion of the N-terminus of the first
helix, which supports the histidine ring. In addition, the C-terminal helix is
bent because the side chain of Leu86 located at the end of this helix partly
fills the hydrophobic cavity created by the mutation. Xenon, which is known to
occupy hydrophobic cavities, causes a partial reversal of the mutation-induced
structural effects. The observed structural changes explain the reduced
phosphocarrier activity of the mutant and agree well with the earlier suggestion
that Ile14 represents an anchoring point stabilizing the active-centre loop in
its correct conformation.
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Figure 4.
Fig. 4. Comparison of wild-type and mutant HPr. Comparison
of the three-dimensional structures of the mutant (left) and
wild-type HPr (right). The side chains of the catalytically
active histidine residue 15 and of residue 14 (isoleucine to
alanine) are shown in blue and yellow, respectively. Residues
Ala19 and Leu86 are indicated in red. The removal of the
isoleucine side chain in the mutant protein leads to significant
structural rearrangements (see text).
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Figure 6.
Fig. 6. Hydrophobic cavity of HPr(I14A). The
solvent-accessible surface of the HPr(I14A) molecule is shown.
Residues 14, 15, 19 and 86 are coloured as in Fig. 4. A cavity
in the region where the mutation has been introduced is marked
by the arrows. The existence of this cavity was confirmed by
xenon-binding studies.
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The above figures are
reprinted
by permission from the Federation of European Biochemical Societies:
Eur J Biochem
(2004,
271,
4815-4824)
copyright 2004.
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Secondary reference #1
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Title
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15n and 1h nmr study of histidine containing protein (hpr) from staphylococcus carnosus at high pressure.
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Authors
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H.R.Kalbitzer,
A.Görler,
H.Li,
P.V.Dubovskii,
W.Hengstenberg,
C.Kowolik,
H.Yamada,
K.Akasaka.
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Ref.
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Protein Sci, 2000,
9,
693-703.
[DOI no: ]
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PubMed id
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Secondary reference #2
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Title
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Solution structure of the histidine-Containing phosphocarrier protein from staphylococcus carnosus
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Authors
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A.Goerler,
W.Hengstenberg,
M.Kravanja,
W.Beneicke,
T.Maurer,
H.R.Kalbitzer.
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Ref.
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appl magn reson, 1999,
17,
465.
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Secondary reference #3
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Title
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Nmr-Spectroscopic mapping of an engineered cavity in the i14a mutant of hpr from staphylococcus carnosus using xenon.
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Authors
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C.Gröger,
A.Möglich,
M.Pons,
B.Koch,
W.Hengstenberg,
H.R.Kalbitzer,
E.Brunner.
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Ref.
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J Am Chem Soc, 2003,
125,
8726-8727.
[DOI no: ]
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PubMed id
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Secondary reference #4
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Title
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Dress: a database of refined solution nmr structures.
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Authors
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S.B.Nabuurs,
A.J.Nederveen,
W.Vranken,
J.F.Doreleijers,
A.M.Bonvin,
G.W.Vuister,
G.Vriend,
C.A.Spronk.
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Ref.
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Proteins, 2004,
55,
483-486.
[DOI no: ]
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PubMed id
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Figure 1.
Figure 1. Distribution of quality scores before and after
refinement. Quality Z-scores distributions of the hundred
selected NMR structures are presented for (A) Ramachandran plot
appearance, (B) 2^nd generation packing quality (all backbone
and side chain contacts), and (C)  [1]-
[2]
rotamer normality. (D) RMS Z-score distribution for heavy atom
side-chain planarity. For a discussion on Z-scores and how to
interpret them, see Linge et al.[17] All distributions are shown
before (red) and after (blue) refinement, values for the WHAT
CHECK reference database are indicated with a dashed line.
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The above figure is
reproduced from the cited reference
with permission from John Wiley & Sons, Inc.
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Secondary reference #5
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Title
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Refinement of protein structures in explicit solvent.
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Authors
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J.P.Linge,
M.A.Williams,
C.A.Spronk,
A.M.Bonvin,
M.Nilges.
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Ref.
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Proteins, 2003,
50,
496-506.
[DOI no: ]
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PubMed id
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Figure 1.
Figure 1. Functional form of the composite cosine term in eq. 1
showing the single maximum at the cis position.
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Figure 2.
Figure 2. Ramachandran map illustrating the aggregate effect of
the four dihedral function for C[  ]-C[
 ]-C-N,
C[ ]-C[
]-C-O,
C-N-C[ ],
and C-N-C[ ]-C
on the energy of the backbone conformation. Contours at 0.4 kcal
mol^-1; intervals with darker shading representing higher energy.
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The above figures are
reproduced from the cited reference
with permission from John Wiley & Sons, Inc.
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