A proteomic approach to apoplastic proteins involved in cell wall regeneration in protoplasts of Arabidopsis suspension-cultured cells

Hye Kyoung Kwon, Ryusuke Yokoyama, Kazuhiko Nishitani

Research output: Contribution to journalArticlepeer-review

102 Citations (Scopus)

Abstract

To clarify the mechanisms of cell wall construction, we used a proteomic approach to investigate the proteins secreted into cell wall spaces during cell wall regeneration from the protoplasts of Arabidopsis suspension-cultured cells. We focused on cell wall proteins loosely bound to the cell wall architecture and extractable with 1 M KCl solutions from: (i) native suspension cultured cells; (ii) protoplasts that had been allowed to regenerate their cell walls for 1 h; and (iii) protoplasts allowed to regenerate their cell walls for 3 h. We adopted a non-destructive extraction procedure without disrupting cellular integrity, thereby avoiding contamination from cytoplasmic proteins. Using two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and matrix-assisted laser desorption ionization-time-of-flight/mass spectrometry (MALDI-TOF/MS), we separated, mapped and identified 71 proteins derived from the native cell wall, and 175 and 212 proteins derived from the 1 and 3 h regenerated protoplasts, respectively. Quite different sets of proteins with differing status of their post-translational modifications, including phosphorylation and glycosylation, were identified in the three protein fractions. This indicated dynamic in muro changes in the cell wall proteins during cell wall regeneration in the protoplasts. The analysis revealed a set of enzymes specifically involved in cell wall expansion and construction in suspension-cultured cells. This approach has also determined a set of cell wall proteins that had not been predicted to be localized in cell wall spaces.

Original languageEnglish
Pages (from-to)843-857
Number of pages15
JournalPlant and Cell Physiology
Volume46
Issue number6
DOIs
Publication statusPublished - 2005 Jun

Keywords

  • Arabidopsis
  • Cell wall regeneration
  • Extracellular proteins
  • Post-translational processing
  • Proteomics
  • Suspension cultured cells

ASJC Scopus subject areas

  • Physiology
  • Plant Science
  • Cell Biology

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