Activity of an atypical Arabidopsis thaliana pectin methylesterase

Planta. 2009 Jan;229(2):311-21. doi: 10.1007/s00425-008-0831-0. Epub 2008 Oct 21.

Abstract

An Arabidopsis thaliana pectin methylesterase that was not predicted to contain any signaling sequence was produced in E. coli and purified using a His tag added at its N-terminus. The enzyme demethylesterified Citrus pectin with a Km of 0.86 mg/ml. The enzyme did not require salt for activity and was found to be relatively temperature-sensitive. The precipitation of enzyme-treated pectin by CaCl2 suggested that the enzyme had a blockwise mode of pectin demethylesterification. A purified kiwi (Actinidia chinensis) pectin methylesterase inhibitor had no effect on the activity of the enzyme whereas it strongly inhibited a flax pectin methylesterase. A model of the protein structure revealed that an extra amino acid sequence in this particular Arabidopsis pectin methylesterase could form a ss-strand outside the core structure, which might be preventing the inhibitor from binding the protein.

MeSH terms

  • Actinidia / enzymology
  • Amino Acid Sequence
  • Arabidopsis / drug effects
  • Arabidopsis / enzymology*
  • Arabidopsis Proteins / antagonists & inhibitors
  • Arabidopsis Proteins / chemistry
  • Arabidopsis Proteins / isolation & purification
  • Arabidopsis Proteins / metabolism*
  • Carboxylic Ester Hydrolases / antagonists & inhibitors
  • Carboxylic Ester Hydrolases / chemistry
  • Carboxylic Ester Hydrolases / isolation & purification
  • Carboxylic Ester Hydrolases / metabolism*
  • Catalytic Domain
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Inhibitors / pharmacology
  • Models, Molecular
  • Molecular Sequence Data
  • Protein Structure, Secondary
  • Sequence Alignment
  • Sequence Analysis, Protein

Substances

  • Arabidopsis Proteins
  • Enzyme Inhibitors
  • Carboxylic Ester Hydrolases
  • pectinesterase