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

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 a...

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Bibliographic Details
Published in:FEBS letters 2017-12, Vol.591 (23), p.3872-3880
Main Authors: Fedosejevs, Eric T., Liu, Leo N.C., Abergel, Megan, She, Yi‐Min, Plaxton, William C.
Format: Article
Language:English
Subjects:
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
protein–protein interactions
Proteomics
reversibly glycosylated polypeptide
Ricinus - chemistry
Ricinus - enzymology
Ricinus - genetics
Ricinus communis - chemistry
Ricinus communis - enzymology
Ricinus communis - genetics
Ricinus communis (castor oil plant)
sucrose synthase
Tandem Mass Spectrometry
UDP‐arabinopyranose mutase
α‐1,4‐glucan protein synthase
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Summary: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.
ISSN:0014-5793
1873-3468
DOI:10.1002/1873-3468.12893