title: Quantification of the brassinosteroid insensitive1 receptor in planta.
abstract: In plants, green fluorescent protein (GFP) is routinely used to determine the subcellular location of fusion proteins. Here, we show that confocal imaging can be employed to approximate the number of GFP-labeled protein molecules present in living Arabidopsis (Arabidopsis thaliana) root cells. The technique involves calibration with soluble GFP to provide a usable protein concentration range within the confocal volume of the microscope. As a proof of principle, we quantified the Brassinosteroid Insensitive1 (BRI1) receptor fused to GFP, under control of its own promoter. The number of BRI1-GFP molecules per root epidermal cell ranges from 22,000 in the meristem and 130,000 in the elongation zone to 80,000 in the maturation zone, indicating that up to 6-fold differences in BRI1 receptor content exist. In contrast, when taking into account differences in cell size, BRI1-GFP receptor density in the plasma membrane is kept constant at 12 receptors μm⁻² in all cells throughout the meristem and elongation zone. Only the quiescent center and columella cells deviate from this pattern and have 5 to 6 receptors μm⁻². Remarkably, root cell sensitivity toward brassinosteroids appears to coincide with uniform meristem receptor density.
authors: Department of Agrotechnology and Food Sciences, Laboratory of Biochemistry, Wageningen University, 6703 HA Wageningen, The Netherlands. wilma.vanesse@wur.nl
authors: G
authors: G
authors: Wilma van Esse
authors: Adrie H
authors: AH
authors: Westphal
authors: Ramya Preethi
authors: RP
authors: Surendran
authors: Catherine
authors: C
authors: Albrecht
authors: Boudewijn
authors: B
authors: van Veen
authors: Jan Willem
authors: JW
authors: Borst
authors: Sacco C
authors: SC
authors: de Vries
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