ArticleJournal of experimental botany2015
A reversible Renilla luciferase protein complementation assay for rapid identification of protein-protein interactions reveals the existence of an interaction network involved in xyloglucan biosynthesis in the plant Golgi apparatus.
Article in Journal of experimental botany, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed, 39 citations in OpenAlex.
- Xyloglucan xylosyltransferase stem region mediates heterodimer formation.Nature communications · 2026Article
- Presence of chronic morbidities alters skeletal muscle health and amino acid kinetics in mild cognitive impairment.Journal of Alzheimer's disease : JAD · 2025Article
- Protein Engineering for Spatiotemporally Resolved Cellular Monitoring.Annual review of analytical chemistry (Palo Alto, Calif.) · 2025Review
- Glucomannan engineering highlights roles of galactosyl modification in fine-tuning cellulose-glucomannan interaction in Arabidopsis cell walls.Nature communications · 2025Article
- Multiprotein Complexes of Plant Glycosyltransferases Involved in Their Function and Trafficking.Plants (Basel, Switzerland) · 2025Review
- Elucidating the molecular logic of a metabotropic glutamate receptor heterodimer.Nature communications · 2024Article
- Article
- Xyloglucan Biosynthesis: From Genes to Proteins and Their Functions.Frontiers in plant science · 2022Review
- Article
- Polysaccharide Biosynthesis: Glycosyltransferases and Their Complexes.Frontiers in plant science · 2021Article
- Interactomes: Experimental and In Silico Approaches.Advances in experimental medicine and biology · 2021Article
- Coelenterazine-Dependent Luciferases as a Powerful Analytical Tool for Research and Biomedical Applications.International journal of molecular sciences · 2020Review
- Pectin Synthesis and Pollen Tube Growth inFrontiers in plant science · 2020Article
- A Hybrid Approach Enabling Large-Scale Glycomic Analysis of Post-Golgi Vesicles Reveals a Transport Route for Polysaccharides.The Plant cell · 2019Article
- Xylan in the Middle: Understanding Xylan Biosynthesis and Its Metabolic Dependencies Toward Improving Wood Fiber for Industrial Processing.Frontiers in plant science · 2019Review
- FRETting about the affinity of bimolecular protein-protein interactions.Protein science : a publication of the Protein Society · 2018Article
- The cell biology of secondary cell wall biosynthesis.Annals of botany · 2018Review
- Enzymatic Activity of Xyloglucan Xylosyltransferase 5.Plant physiology · 2016Article
- Techniques for the Analysis of Protein-Protein Interactions in Vivo.Plant physiology · 2016Review
- Asparagus IRX9, IRX10, and IRX14A Are Components of an Active Xylan Backbone Synthase Complex that Forms in the Golgi Apparatus.Plant physiology · 2016Article
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Authors and funding
6 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A growing body of evidence suggests that protein-protein interactions (PPIs) occur amongst glycosyltransferases (GTs) required for plant glycan biosynthesis (e.g. cell wall polysaccharides and N-glycans) in the Golgi apparatus, and may control the functions of these enzymes. However, identification of PPIs in the endomembrane system in a relatively fast and simple fashion is technically challenging, hampering the progress in understanding the functional coordination of the enzymes in Golgi glycan biosynthesis. To solve the challenges, we adapted and streamlined a reversible Renilla luciferase protein complementation assay (Rluc-PCA), originally reported for use in human cells, for transient expression in Nicotiana benthamiana. We tested Rluc-PCA and successfully identified luminescence complementation amongst Golgi-localizing GTs known to form a heterodimer (GAUT1 and GAUT7) and those which homooligomerize (ARAD1). In contrast, no interaction was shown between negative controls (e.g. GAUT7, ARAD1, IRX9). Rluc-PCA was used to investigate PPIs amongst Golgi-localizing GTs involved in biosynthesis of hemicelluloses. Although no PPI was identified among six GTs involved in xylan biosynthesis, Rluc-PCA confirmed three previously proposed interactions and identified seven novel PPIs amongst GTs involved in xyloglucan biosynthesis. Notably, three of the novel PPIs were confirmed by a yeast-based split-ubiquitin assay. Finally, Gateway-enabled expression vectors were generated, allowing rapid construction of fusion proteins to the Rluc reporters and epitope tags. Our results show that Rluc-PCA coupled with transient expression in N. benthamiana is a fast and versatile method suitable for analysis of PPIs between Golgi resident proteins in an easy and mid-throughput fashion in planta.
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