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Lateral diffusion of proteins and lipids in the plasma membrane of rose protoplast

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Summary

Lateral diffusion measurements have been made on lipids and proteins in the plasma membrane of live protoplasts derived from rose (Rosa sp. “Paul's Scarlet”) suspension-cultured cells. Two different fluorescent lipid probes exhibited markedly different diffusion rates, indicating possible heterogeneity in the lipid domain of the membrane. Membrane proteins were labeled directly with covalently-reactive fluorophores, and factors that might perturb the lateral diffusion of these labeled proteins were investigated. Treatment of the protoplasts with various cytoskeleton-disrupting drugs generally had little effect on protein diffusion, although treatment with oryzalin, a microtubule-disrupting drug, did slightly reduce the mobile fraction of membrane proteins. Elevation of the CaCl2 concentration in the medium from 1 mM to 10 mM significantly reduced the mobile fraction of membrane proteins and also increased the fraction of protoplasts that were able to regenerate cell walls and divide in culture. These results are discussed in relation to reported evidence of lipid domains in the plasma membranes of other cells and protoplasts. The relative importance of lipid domains and membrane-cytoskeleton interaction in governing protein diffusion is considered.

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Abbreviations

D:

lateral diffusion coefficient

RCA:

Ricinus communis agglutinin

BPA:

Bauhinia purpurea agglutinin

DTAF:

dichlorotriazinylaminofluorescein

FTSC:

fluorescein-5-thiosemicarbazide

C18-Fl:

5-(N-octadecanoyl)aminofluorescein

LY-Chol:

Lucifer yellow conjugate of cholesterol, i.e., dilithium 4-amino-N-[(β-(carbo(5-cho-lesten-3β-yl)oxy)hydrazinocarbonyl)amino]-1,8-naphthalimide-3,6-disulfonate

APM:

amiprophosmethyl

DMSO:

dimethylsulfoxide

FPR:

fluorescence photobleaching recovery

sd:

standard deviation

FRAF:

fluorescence redistribution after fusion

M:

mobile fraction

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Author notes

  1. R. M. Walko

    Present address: USDA Plant Gene Expression Center, 800 Buchanan Street, 94710, Albany, CA, USA

Authors and Affiliations

  1. Department of Botany and Plant Sciences, University of California, Riverside, California

    R. M. Walko & E. A. Nothnagel

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  1. R. M. Walko
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  2. E. A. Nothnagel
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Walko, R.M., Nothnagel, E.A. Lateral diffusion of proteins and lipids in the plasma membrane of rose protoplast. Protoplasma 152, 46–56 (1989). https://doi.org/10.1007/BF01354239

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  • DOI: https://doi.org/10.1007/BF01354239

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