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PDBsum entry 1j0r
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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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The impact of single cysteine residue mutations on the replication terminator protein.
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Authors
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J.P.Vivian,
A.F.Hastings,
I.G.Duggin,
R.G.Wake,
M.C.Wilce,
J.A.Wilce.
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Ref.
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Biochem Biophys Res Commun, 2003,
310,
1096-1103.
[DOI no: ]
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PubMed id
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Abstract
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We report the structural and biophysical consequences of cysteine substitutions
in the DNA-binding replication terminator protein (RTP) of Bacillus subtilis,
that resulted in an optimised RTP mutant suitable for structural studies. The
cysteine residue 110 was replaced with alanine, valine or serine. Protein
secondary structure and stability (using circular dichroism spectropolarimetry),
self-association (using analytical ultracentrifugation), and DNA-binding
measurements revealed RTP.C110S to be the most similar mutant to wild-type RTP.
The C110A and C110V.RTP mutants were less soluble, less stable and showed lower
DNA-binding affinity. The structure of RTP.C110S, solved to 2.5A resolution
using crystallographic methods, showed no major structural perturbation due to
the mutation. Heteronuclear NMR spectroscopic studies revealed subtle
differences in the electronic environment about the site of mutation. The study
demonstrates the suitability of serine as a substitute for cysteine in RTP and
the high sensitivity of protein behaviour to single amino acid substitutions.
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