Determination of structural regions important for Ca 2+ uptake activity in arabidopsis MCA1 and MCA2 expressed in yeast

Masataka Nakano, Kazuko Iida, Hiroshi Nyunoya, Hidetoshi Iida

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

MCA1 is a plasma membrane protein that correlates Ca 2+ influx and mechanosensing in Arabidopsis. MCA2 is a paralog of MCA1, and both share 72.7 amino acid sequence identity and several common structural features, including putative transmembrane (TM) segments, an EF hand-like region in the N-terminal half, a coiled-coil motif in the middle and a PLAC8 motif in the C-terminal half. To determine structural regions important for Ca 2+ uptake activity, the activity of truncated forms of MCA1 and MCA2 was assessed using yeast expression assays. The N-terminal half of MCA1 with a coiled-coil motif (MCA1 1-237) did not have Ca 2+ uptake activity, while MCA2 1-237 did. The N-terminal half of MCA1 without the coiled-coil motif (MCA1 1-185) showed Ca 2+ uptake activity, as did MCA2 1-186. Both MCA1 1-173 and MCA2 1-173 having the EF hand-like region had Ca 2+ uptake activity. Deletion of a putative TM segment (Ile11-Ala33) and the Asp21 to asparagine mutation in MCA1 and MCA2 abolished Ca 2+ uptake activity. Finally, MCA1 173-421 and MCA2 173-416 lacking the N-terminal half had no Ca 2+ uptake activity. These results suggest that the N-terminal half of both proteins with the EF hand-like region is necessary and sufficient for Ca 2+ uptake and that the coiled-coil motif regulates MCA1 negatively and MCA2 positively.

Original languageEnglish
Pages (from-to)1915-1930
Number of pages16
JournalPlant and Cell Physiology
Volume52
Issue number11
DOIs
Publication statusPublished - 2011 Nov
Externally publishedYes

Fingerprint

EF Hand Motifs
Arabidopsis
Yeasts
yeasts
uptake mechanisms
Asparagine
hands
Blood Proteins
Amino Acid Sequence
Membrane Proteins
Cell Membrane
Mutation
asparagine
membrane proteins
Proteins
blood proteins
plasma membrane
amino acid sequences
mutation
assays

Keywords

  • Arabidopsis thaliana
  • Ca channel
  • Ca transport
  • Heterologous expression
  • Saccharomyces cerevisiae
  • Structure-function relationship

ASJC Scopus subject areas

  • Physiology
  • Plant Science
  • Cell Biology

Cite this

Determination of structural regions important for Ca 2+ uptake activity in arabidopsis MCA1 and MCA2 expressed in yeast. / Nakano, Masataka; Iida, Kazuko; Nyunoya, Hiroshi; Iida, Hidetoshi.

In: Plant and Cell Physiology, Vol. 52, No. 11, 11.2011, p. 1915-1930.

Research output: Contribution to journalArticle

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