PDBsum entry 1j0r

Replication

PDB id

1j0r

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Contents

			Protein chains

					114 a.a.

			Waters ×58

PDB id:

1j0r

Links

Name:

Replication

Title:

Crystal structure of the replication termination protein mutant c110s

Structure:

Replication termination protein. Chain: a, b. Synonym: replication terminator protein. Engineered: yes. Mutation: yes

Source:

Bacillus subtilis. Organism_taxid: 1423. Expressed in: escherichia coli. Expression_system_taxid: 562.

Biol. unit:

Dimer (from PDB file)

Resolution:

2.50Å

R-factor:

0.233

R-free:

0.279

Authors:

J.P.Vivian,A.F.Hastings,I.G.Duggin,R.G.Wake,M.C.J.Wilce,J.A.Wilce

Key ref:

J.P.Vivian et al. (2003). The impact of single cysteine residue mutations on the replication terminator protein. Biochem Biophys Res Commun, 310, 1096-1103. PubMed id: 14559228 DOI: 10.1016/j.bbrc.2003.09.126

Date:

20-Nov-02

Release date:

20-Nov-03

PROCHECK

	Headers

	References

Protein chains

P0CI76 (RTP_BACSU) - Replication termination protein from Bacillus subtilis (strain 168)

Seq: Struc:					122 a.a. 114 a.a.*

Key:

PfamA domain

Secondary structure

CATH domain

* PDB and UniProt seqs differ at 1 residue position (black cross)

DOI no: 10.1016/j.bbrc.2003.09.126	Biochem Biophys Res Commun 310:1096-1103 (2003)
PubMed id: 14559228

The impact of single cysteine residue mutations on the replication terminator protein.
J.P.Vivian, A.F.Hastings, I.G.Duggin, R.G.Wake, M.C.Wilce, J.A.Wilce.

ABSTRACT

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.

Literature references that cite this PDB file's key reference

PubMed id

Reference

17077489

J.P.Vivian, C.Porter, J.A.Wilce, and M.C.Wilce (2006).
Crystallization and preliminary X-ray diffraction analysis of the Bacillus subtilis replication termination protein in complex with the 37-base-pair TerI-binding site.

Acta Crystallogr Sect F Struct Biol Cryst Commun, 62, 1104-1107.

15841400

C.J.Porter, M.C.Wilce, J.P.Mackay, P.Leedman, and J.A.Wilce (2005).
Grb7-SH2 domain dimerisation is affected by a single point mutation.

Eur Biophys J, 34, 454-460.

The most recent references are shown first. Citation data come partly from CiteXplore and partly from an automated harvesting procedure. Note that this is likely to be only a partial list as not all journals are covered by either method. However, we are continually building up the citation data so more and more references will be included with time.