Part:BBa_K808003

tctB_162: small subunit B1 of the tripartite tricarboxylate transporter family

The small subunit B1 of the tripartite tricarboxylate transporter family (tctB_162, 17 kDa) was isolated from Comamonas testosteroni KF-1. The tripartite tricarboxylate transporter system consists of three different proteins: a periplasmatic solute binding receptor, a membrane protein with 12 putative transmembrane alpha-helical spanners (in this case tctB_162), and a small poorly conserved membrane proteine with four putative transmembrane alpha-helical spanners^[1].The strain was purchased from Leibniz Institute DMSZ-German Collection of Microorganism and Cell Cultures (DMSZ no. 14576). The original sequence contains a Pst1 recognition site. To eliminate this recognition site a directed-site mutagenic PCR was performed. (For more datails: link zu dem PCR protokoll und dem Labjournal, wo isolations PCR beschrieben wird) To characterized the structure of the tctB_162 bioinformatic tools like P rotein H omology/anolog Y R ecognition E ngine V 2.0 (PHYRE2), I-TASSER servers, protein B asic L ocal A ligment S earch T ool (BLAST) and TMHMM was used. The TMHMM predicted a transmembrane protein with 5 alpha-helical spanners (Fig. 1). The N-teminus is with a probability of over 99 % in cytoplasmatic. The NCBI Protein BLAST results shows that the tctB_162 subunit B1 belongs to the tctB superfamily.

Figure 1. TMHMM prediction of the tctB_162 subunit B1 It shows 5 alpha-helical spanners.

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 NgoMIV site found at 291
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 55
  

References

[1] Sasoh, M., E. Masai, et al. (2006). "Characterization of the terephthalate degradation genes of Comamonas sp. strain E6." Appl Environ Microbiol 72(3): 1825-1832.