Abstract
The endoplasmic reticulum takes care of the folding, assembly, and quality control of thousands of proteins destined to the different compartments of the endomembrane system, or to be secreted in the apoplast. Here we describe how these early events in the life of all these proteins can be followed biochemically by using velocity or isopycnic ultracentrifugation, metabolic labeling with radioactive amino acids, and immunoprecipitation in various conditions.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Vitale A, Ceriotti A, Denecke J (1993) The role of the endoplasmic reticulum in protein synthesis modification and intracellular transport. J Exp Bot 44:1417–1444
Vitale A, Boston R (2008) Endoplasmic reticulum quality control and the unfolded protein response: insights from plants. Traffic 9:1581–1588
Liu XJ, Howell SH (2016) Managing the protein folding demands in the endoplasmic reticulum of plants. New Phytol 211:418–428
Zhang F, Boston RS (1992) Increases in binding protein (BiP) accompany changes in protein body morphology in three high lysine mutants of maize. Protoplasma 171:142–152
Pedrazzini E, Giovinazzo G, Bollini R, Ceriotti A, Vitale A (1994) Binding of BiP to an assembly-defective protein in plant cells. Plant J 5:103–110
Pedrazzini E, Giovinazzo G, Bielli A, de Virgilio M, Frigerio L, Pesca M, Faoro F, Bollini R, Ceriotti A, Vitale A (1997) Protein quality control along the route to the plant vacuole. Plant Cell 9:1869–1880
Hong Z, Jin H, Tzfira T, Li J (2008) Multiple mechanism-mediated retention of a defective brassinosteroid receptor in the endoplasmic reticulum of Arabidopsis. Plant Cell 20:3418–3429
Maîtrejean M, Wudick MM, Voelker C, Prinsi B, Mueller-Roeber B, Czempinski K, Pedrazzini E, Vitale A (2011) Assembly and sorting of the tonoplast potassium channel AtTPK1 and its turnover by internalization into the vacuole. Plant Physiol 156:1783–1796
Pollard JK, Steward FC (1959) The use of proline-C14 by growing cells: its conversion to protein and hydroxyproline. J Exp Bot 10:17–32
Ceresoli V, Mainieri D, Del Fabbro M, Weinstein R, Pedrazzini E (2016) A fusion between domains of the human bone morphogenetic protein-2 and maize 27 kD γZein accumulates to high levels in the endoplasmic reticulum without forming protein bodies in transgenic tobacco. Front Plant Sci 7:358
Barbante A, Irons S, Hawes C, Frigerio L, Vitale A, Pedrazzini E (2008) Anchorage to the cytosolic face of the endoplasmic reticulum membrane: a new strategy to stabilize a cytosolic recombinant antigen in plants. Plant Biotechnol J 6:560–575
Pedrazzini E, Mainieri D, Marrano CA, Vitale A (2016) Where do protein bodies of cereal seeds come from? Front Plant Sci 7:1139
Shani N, Rosenberg N, Kasarda DD, Galili G (1994) Mechanisms of assembly of wheat high molecular weight glutenins inferred from expression of wild-type and mutant subunits in transgenic tobacco. J Biol Chem 269:8924–8930
Mainieri D, Rossi M, Archinti M, Bellucci M, De Marchis F, Vavassori S, Pompa A, Arcioni S, Vitale A (2004) Zeolin, a new recombinant storage protein constructed using maize gamma-zein and bean phaseolin. Plant Physiol 136:3447–3456
Pompa A, Vitale A (2006) Retention of a bean phaseolin/maize γ-zein fusion in the endoplasmic reticulum depends on disulfide bond formation. Plant Cell 18:2608–2621
Foresti O, Frigerio L, Holkeri H, de Virgilio M, Vavassori S, Vitale A (2003) A phaseolin domain directly involved in trimer assembly is a BiP binding determinant. Plant Cell 15:2464–2475
Acknowledgements
We thank the present and past members of our laboratories for their fundamental contributions in developing and continuously improving the methods described here.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Pedrazzini, E., Vitale, A. (2018). Protein Biosynthesis and Maturation in the ER. In: Hawes, C., Kriechbaumer, V. (eds) The Plant Endoplasmic Reticulum . Methods in Molecular Biology, vol 1691. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7389-7_14
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7389-7_14
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7388-0
Online ISBN: 978-1-4939-7389-7
eBook Packages: Springer Protocols