Journal:

Article Title: Bacterial chromosome segregation: structure and DNA binding of the Soj dimer -- a conserved biological switch

doi: 10.1038/sj.emboj.7600530

Figure Lengend Snippet: Size exclusion chromatography of T. thermophilus wild-type Soj and Soj D44A in the presence and absence of ATP. The elution volume of each species is indicated in millilitres above the respective trace. (A) In the absence of ATP, wild-type Soj elutes as a monomeric species (black). In the presence of ATP, Soj elutes as a dimer–monomer equilibrium (green). (B) Soj D44A, deficient in nucleotide hydrolysis, also elutes as a monomer in the absence of ATP (black), but elutes almost solely as a dimer in the presence of ATP (green). Red: absorbance at 260 nm indicating the elution volume of ATP and the presence of ATP in the dimeric protein.

Article Snippet: Soj from T. thermophilus HB27 (ATCC BAA-163D) was cloned into pHis17 to generate a C-terminally hexa-histidine-tagged protein.

Techniques: Size-exclusion Chromatography

Journal:

Article Title: Bacterial chromosome segregation: structure and DNA binding of the Soj dimer -- a conserved biological switch

doi: 10.1038/sj.emboj.7600530

Figure Lengend Snippet: Sequence alignment of putative activating peptides of Spo0J, MinE and FtsA. Alignment of the N-terminal regions of Spo0J and MinE proteins with the C-terminus of FtsA proteins reveals a remarkable sequence homology, which includes a universally conserved basic (lys/arg) residue (arginine 10 in T. thermophilus Spo0J). The functional homology of these putative activating peptides is further strengthened by the observations that E. coli FtsA C-terminus can stimulate Soj and mutation of the arginine 10 to alanine in the Spo0JN20 peptide abrogates stimulation of ATP hydrolysis (Figure 7).

Article Snippet: Soj from T. thermophilus HB27 (ATCC BAA-163D) was cloned into pHis17 to generate a C-terminally hexa-histidine-tagged protein.

Techniques: Sequencing, Functional Assay, Mutagenesis