A novel TctA citrate transporter from an activated sludge metagenome: structural and mechanistic predictions for the TTT family

Proteins. 2014 Sep;82(9):1756-64. doi: 10.1002/prot.24529. Epub 2014 Feb 19.

Abstract

We isolated a putative citrate transporter of the tripartite tricarboxylate transporter (TTT) class from a metagenomic library of activated sludge from a sewage treatment plant. The transporter, dubbed TctA_ar, shares ∼50% sequence identity with TctA of Comamonas testosteroni (TctA_ct) and other β-Proteobacteria, and contains two 20-amino acid repeat signature sequences, considered a hallmark of this particular transporter class. The structures for both TctA_ar and TctA_ct were modeled with I-TASSER and two possible structures for this transporter family were proposed. Docking assays with citrate resulted in the corresponding sets of proposed critical residues for function. These models suggest functions for the 20-amino acid repeats in the context of the two different architectures. This constitutes the first attempt at structure modeling of the TTT family, to the best of our knowledge, and could aid functional understanding of this little-studied family.

Keywords: TctA; citrate; docking; gene phylogeny; metagenomic; molecular modeling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Binding Sites
  • Carrier Proteins / chemistry*
  • Carrier Proteins / isolation & purification
  • Carrier Proteins / ultrastructure*
  • Citric Acid / chemistry
  • Comamonas testosteroni / enzymology*
  • Comamonas testosteroni / genetics
  • Gene Library
  • Metagenome / genetics
  • Molecular Docking Simulation
  • Molecular Sequence Data
  • Protein Binding
  • Protein Structure, Tertiary
  • Repetitive Sequences, Amino Acid / genetics*
  • Sequence Alignment
  • Sewage / microbiology

Substances

  • Carrier Proteins
  • Sewage
  • citrate-binding transport protein
  • Citric Acid