An extracellular hydrophilic carboxy-terminal domain regulates the activity of TaALMT1, the aluminum-activated malate transport protein of wheat

Takuya Furuichi, Takayuki Sasaki, Yoshiyuki Tsuchiya, Peter R. Ryan, Emmanuel Delhaize, Yoko Yamamoto

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Al3+-resistant cultivars of wheat (Triticum aestivum L.) release malate through the Al3+-activated anion transport protein Triticum aestivum aluminum-activated malate transporter 1 (TaALMT1). Expression of TaALMT1 in Xenopus oocytes and tobacco suspension cells enhances the basal transport activity (inward and outward currents present in the absence of external Al3+), and generates the same Al3+-activated currents (reflecting the Al3+-dependent transport function) as observed in wheat cells. We investigated the amino acid residues involved in this Al3+-dependent transport activity by generating a series of mutations to the TaALMT1 protein. We targeted the acidic residues on the hydrophilic C-terminal domain of TaALMT1 and changed them to uncharged residues by site-directed mutagenesis. These mutant proteins were expressed in Xenopus oocytes and their transport activity was measured before and after Al 3+ addition. Three mutations (E274Q, D275N and E284Q) abolished the Al3+-activated transport activity without affecting the basal transport activity. Truncation of the hydrophilic C-terminal domain abolished both basal and Al3+-activated transport activities. Al 3+-dependent transport activity was recovered by fusing the N-terminal region of TaALMT1 with the C-terminal region of AtALMT1, a homolog from Arabidopsis. These findings demonstrate that the extracellular C-terminal domain is required for both basal and Al3+-dependent TaALMT1 activity. Furthermore, we identified three acidic amino acids within this domain that are specifically required for the activation of transport function by external Al3+.

Original languageEnglish
Pages (from-to)47-55
Number of pages9
JournalPlant Journal
Volume64
Issue number1
DOIs
Publication statusPublished - Oct 2010

Fingerprint

transport proteins
malates
Aluminum
Triticum
aluminum
transporters
Carrier Proteins
Triticum aestivum
wheat
Xenopus
Oocytes
Anion Transport Proteins
Acidic Amino Acids
malic acid
Mutation
Mutant Proteins
Site-Directed Mutagenesis
Arabidopsis
oocytes
Tobacco

Keywords

  • acidic amino acids
  • aluminum activation
  • malate transport
  • Triticum aestivum aluminum-activated malate transporter 1
  • two-electrode voltage clamp
  • Xenopus oocyte

ASJC Scopus subject areas

  • Plant Science
  • Cell Biology
  • Genetics

Cite this

An extracellular hydrophilic carboxy-terminal domain regulates the activity of TaALMT1, the aluminum-activated malate transport protein of wheat. / Furuichi, Takuya; Sasaki, Takayuki; Tsuchiya, Yoshiyuki; Ryan, Peter R.; Delhaize, Emmanuel; Yamamoto, Yoko.

In: Plant Journal, Vol. 64, No. 1, 10.2010, p. 47-55.

Research output: Contribution to journalArticle

Furuichi, Takuya ; Sasaki, Takayuki ; Tsuchiya, Yoshiyuki ; Ryan, Peter R. ; Delhaize, Emmanuel ; Yamamoto, Yoko. / An extracellular hydrophilic carboxy-terminal domain regulates the activity of TaALMT1, the aluminum-activated malate transport protein of wheat. In: Plant Journal. 2010 ; Vol. 64, No. 1. pp. 47-55.
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