Part:BBa_K352008

H77T mutated CooA from Rhodospirillum rubrum

This part is the mutated version of CooA protein coding sequence. The mutation involves conversion histidine to threonine at 77th amino acid. The binding strength of pCooF to CooA is expected to diminish.

CooA transcriptional activator makes possible to sense CO and use CO as single energy source for Rhodospirillum rubrum and also it has heme groups.

CooA founds under a transcriptional regulators family which resembles the cAMP receptor protein and fumavate nitrate reduction from Escherichia coli. The protein functions upon in sequence-specific DNA binding when CO exists at the environment. CO dependent CooA is at the dimer structure when CO does not exist. CooA, is 28% identical (51% similar) to CRP(cAMP receptor protein) and 18% identical (45% similar) to FNR(fumavate nitrate reduction) of the Escherichia coli.

The strain has 1FT9 ID number from PDB was used for adaptation of the inactive Fe(II) CooA structure. Two monomers of the protein darkened distinctively. These monomers dimerize along the middle C-helices of adjacent effector-binding domains. Identified structure of the protein is not symmetric and one monomer has fused C- and D-helices. However, two F-helices are away from the surface structure and they act on DNA in a sequence-specific manner. The 4/5 loop, the Pro2 and His77 heme Fe(II) ligands are noted.

Terminator site was added to downstream region the gene.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 466
  
• 1000
  COMPATIBLE WITH RFC[1000]