Part:BBa_K808002

tctA_503: large subunit A1 of the tripartite tricarboxylate transporter family

The large subunit A1 of the tripartite tricarboxylate transporter family (tctA_503, 52,98 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(in this case tctA_503), and a small poorly conserved membrane proteine with four putative transmembrane alpha-helical spanner^[1]. The strain was purchased from 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:[http://2012.igem.org/Team:TU_Darmstadt/Protocols/mutagenic_PCR mutagenic PCR]) To characterized the structure of the tctA_503 bioinformatic tools like Protein Homology/anologY Recognition Engine V 2.0 (PHYRE2), I-TASSER servers, and TMHMM was used. The TMHMM predicted a transmembrane protein with 11 alpha-helical spanners (Fig. 1). The N-teminus is with a probability of 98 % in cytoplasmatic. The NCBI Protein BLAST results shows that the tctA_503 subunit A1 belongs to the tctA superfamily. PHYRE2 and I-Tasser server homology modelling did not give a significant result for the structure of the tctA_503 subunit A1.

Figure 1. TMHMM prediction of the tctA_503 subunit A1. It shows 11-12 transmembrane alpha-helics.

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 1467
  Illegal AgeI site found at 712
  
• 1000
  COMPATIBLE WITH RFC[1000]

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.

• Fukuhara, Y., K. Inakazu, et al. (2010). "Characterization of the isophthalate degradation genes of Comamonas sp. strain E6." Appl Environ Microbiol 76(2): 519-527.
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• Winnen, B., R. N. Hvorup, et al. (2003). "The tripartite tricarboxylate transporter (TTT) family." Res Microbiol 154(7): 457-465.
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