Biochemical characterization of rhamnosyltransferase involved in biosynthesis of pectic rhamnogalacturonan I in plant cell wall

Biochem Biophys Res Commun. 2017 Apr 22;486(1):130-136. doi: 10.1016/j.bbrc.2017.03.012. Epub 2017 Mar 8.

Abstract

The pectin in plant cell walls consists of three domains: homogalacturonan, rhamnogalacturonan (RG)-I, and RG-II. It is predicted that around 50 different glycosyltransferases are required for their biosynthesis. Among these, the activities of only a few glycosyltransferases have been detected because pectic oligosaccharides are not readily available for use as substrates. In this study, fluorogenic pyridylaminated RG-I-backbone oligosaccharides (PA-RGs) with 3-14 degrees of polymerization (DP) were prepared. Using these oligosaccharides, the activity of RG-I:rhamnosyltransferase (RRT), involved in the biosynthesis of the RG-I backbone diglycosyl repeating units (-4GalUAα1-2Rhaα1-), was detected from the microsomes of azuki bean epicotyls. RRT was found to prefer longer acceptor substrates, PA-RGs with a DP > 7, and it does not require any metal ions for its activity. RRT is located in the Golgi and endoplasmic reticulum. The activity of RRT coincided with epicotyl growth, suggesting that RG-I biosynthesis is involved in plant growth.

Keywords: Fluorescent labeling; Glycosyltransferase; Pectin; Plant cell wall; Rhamnogalacturonan I; Rhamnosyltransferase.

MeSH terms

  • Biocatalysis
  • Cell Wall / enzymology
  • Cell Wall / metabolism*
  • Chromatography, High Pressure Liquid
  • Endoplasmic Reticulum / enzymology
  • Endoplasmic Reticulum / metabolism
  • Glycosyltransferases / metabolism*
  • Golgi Apparatus / enzymology
  • Golgi Apparatus / metabolism
  • Magnetic Resonance Spectroscopy
  • Oligosaccharides / metabolism
  • Pectins / biosynthesis*
  • Plant Proteins / metabolism*
  • Substrate Specificity
  • Tandem Mass Spectrometry
  • Vigna / enzymology
  • Vigna / metabolism

Substances

  • Oligosaccharides
  • Plant Proteins
  • rhamnogalacturonan I
  • Pectins
  • Glycosyltransferases