Coimmunoprecipitation of reversibly glycosylated polypeptide with sucrose synthase from developing castor oilseeds

FEBS Lett. 2017 Dec;591(23):3872-3880. doi: 10.1002/1873-3468.12893. Epub 2017 Nov 14.

Abstract

The sucrose synthase (SUS) interactome of developing castor oilseeds (COS; Ricinus communis) was assessed using coimmunoprecipitation (co-IP) with anti-(COS RcSUS1)-IgG followed by proteomic analysis. A 41-kDa polypeptide (p41) that coimmunoprecipitated with RcSUS1 from COS extracts was identified as reversibly glycosylated polypeptide-1 (RcRGP1) by LC-MS/MS and anti-RcRGP1 immunoblotting. Reciprocal Far-western immunodot blotting corroborated the specific interaction between RcSUS1 and RcRGP1. Co-IP using anti-(COS RcSUS1)-IgG and clarified extracts from other developing seeds as well as cluster (proteoid) roots of white lupin and Harsh Hakea consistently recovered 90 kDa SUS polypeptides along with p41/RGP as a SUS interactor. The results suggest that SUS interacts with RGP in diverse sink tissues to channel UDP-glucose derived from imported sucrose into hemicellulose and/or glycoprotein/glycolipid biosynthesis.

Keywords: Ricinus communis (castor oil plant); UDP-arabinopyranose mutase; protein-protein interactions; reversibly glycosylated polypeptide; sucrose synthase; α-1,4-glucan protein synthase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Far-Western
  • Glucosyltransferases / chemistry
  • Glucosyltransferases / genetics
  • Glucosyltransferases / isolation & purification*
  • Glycoproteins / chemistry
  • Glycoproteins / genetics
  • Glycoproteins / isolation & purification
  • Glycosylation
  • Immunoprecipitation
  • Phylogeny
  • Plant Proteins / chemistry
  • Plant Proteins / genetics
  • Plant Proteins / isolation & purification*
  • Protein Interaction Mapping
  • Proteomics
  • Ricinus / chemistry*
  • Ricinus / enzymology*
  • Ricinus / genetics
  • Ricinus communis / chemistry*
  • Ricinus communis / enzymology*
  • Ricinus communis / genetics
  • Tandem Mass Spectrometry

Substances

  • Glycoproteins
  • Plant Proteins
  • Glucosyltransferases
  • sucrose synthase

Associated data

  • GENBANK/XP_002514626
  • GENBANK/XP_002512057
  • GENBANK/XP_002511153
  • GENBANK/XP_002525910
  • GENBANK/XP_002512611