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PDBsum entry 1pfh
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Transport protein
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PDB id
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1pfh
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References listed in PDB file
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Key reference
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Title
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High-Resolution structure of the phosphorylated form of the histidine-Containing phosphocarrier protein hpr from escherichia coli determined by restrained molecular dynamics from nmr-Noe data.
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Authors
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N.A.Van nuland,
R.Boelens,
R.M.Scheek,
G.T.Robillard.
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Ref.
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J Mol Biol, 1995,
246,
180-193.
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PubMed id
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Abstract
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The solution structure of the phosphorylated form of the histidine-containing
phosphocarrier protein, HPr, from Escherichia coli has been determined by NMR in
combination with restrained molecular dynamics simulations. The structure of
phospho-HPr (P-HPr) results from a molecular dynamics simulation in water, using
time-dependent distance restraints to attain agreement with the measured NOEs.
Experimental restraints were identified from both three-dimensional 1H-1H-15N
HSQC-NOESY and two-dimensional 1H-1HNOESY spectra, and compared with those of
the unphosphorylated form. Structural changes upon phosphorylation of HPr are
limited to the active site, as evidenced by changes in chemical shifts, in 3JNHH
alpha-coupling constants and NOE patterns. Chemical shift changes were obtained
mainly for protons that were positioned close to the phosphoryl group attached
to the His15 imidazole ring. Differences could be detected in the intensity of
the NOEs involving the side-chain protons of His15 and Pro18, resulting from a
change in the relative position of the two rings. In addition, a small change
could be detected in the three-bond J-coupling between the amide proton and the
H alpha proton of Thr16 and Arg17 upon phosphorylation, in agreement with the
changes of the phi torsion angle of these two residues obtained from
time-averaged restrained molecular dynamics simulations in water. The proposed
role of the torsion-angle strain at residue 16 in the mechanism of Streptococcus
faecalis HPr is not supported by these results. In contrast, phosphorylation
seems to introduce torsion angle strain at residue His15. This strain could
facilitate the transfer of the phosphoryl group to the A-domain at enzyme II.
The phospho-histidine is not stabilised by hydrogen bonds to the side-chain
group of Arg17; instead stable hydrogen bonds are formed between the phosphate
group and the backbone amide protons of Thr16 and Arg17, which show the largest
changes in chemical shift upon phosphorylation, and a hydrogen bond involving
the side-chain O gamma proton of Thr16. HPr accepts the phosphoryl group from
enzyme I and donates it subsequently to the A domain of various enzyme II
species. The binding site for EI on HPr resembles that of the A domain of the
mannitol-specific enzyme II, as can be concluded from the changes on the amide
proton and nitrogen chemical shifts observed via heteromolecular single-quantum
coherence spectroscopy.
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Secondary reference #1
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Title
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The high-Resolution structure of the histidine-Containing phosphocarrier protein hpr from escherichia coli determined by restrained molecular dynamics from nuclear magnetic resonance nuclear overhauser effect data.
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Authors
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N.A.Van nuland,
I.W.Hangyi,
R.C.Van schaik,
H.J.Berendsen,
W.F.Van gunsteren,
R.M.Scheek,
G.T.Robillard.
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Ref.
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J Mol Biol, 1994,
237,
544-559.
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PubMed id
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Secondary reference #2
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Title
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Active-Centre torsion-Angle strain revealed in 1.6 a-Resolution structure of histidine-Containing phosphocarrier protein.
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Authors
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Z.Jia,
M.Vandonselaar,
J.W.Quail,
L.T.Delbaere.
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Ref.
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Nature, 1993,
361,
94-97.
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PubMed id
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