Measuring plant protein secretion

Larson, E. R. (2017) Measuring plant protein secretion. In: Jiang, L. (ed.) Plant Protein Secretion: Methods and Protocols. Series: Methods in molecular biology (1662). Humana Press, pp. 199-207. ISBN 9781493972616 (doi: 10.1007/978-1-4939-7262-3_18)

Abstract

Fluorescent protein fusions are widely used for visualizing the subcellular localization and mobility of intercellular proteins. There is now a variety of colors, expression vectors, and photoactivated molecules to choose from, each with their own strengths and limitations. In this chapter, the methodologies for expressing and quantifying protein secretion with fluorescent protein fusion constructs using two separate protocols-one in which the retention of a transiently expressed fluorescent marker is measured in seedling roots to quantify a block in secretion, and one in which the secretion of a fluorescent marker into the space of the apoplast is measured to quantify secretion in plant leaves-are described. In the first protocol, seedling roots are transiently transformed with multicistronic constructs; and in the second protocol, markers can be stably expressed and controlled under an inducible promoter in mature plants. Both methods provide tools for quantifying protein secretion and visualizing defects in secretion pathways in Arabidopsis.

• Item Type: Book Sections
• Keywords: Confocal microscopy, fluorescence quantification, multicistronic expression vectors, optobiology, protein secretion, transient expression.
• Status: Published
• Glasgow Author(s) Enlighten ID: Larson, Dr Emily
• Authors: Larson, E. R.
• College/School: College of Medical Veterinary and Life Sciences > School of Molecular Biosciences
• Journal Name: Methods in molecular biology (Clifton, N.J.)
• Publisher: Humana Press
• ISSN: 1940-6029
• ISBN: 9781493972616
• Published Online: 01 September 2017