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

Walko, R. M.; Nothnagel, Eugene A.

doi:10.1007/bf01354239

Cited by 16 publications

(9 citation statements)

References 47 publications

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“…Fluorescence photobleaching recovery experiments carried out in our laboratory (3,4) have yielded results that are comparable to those reported by Metcalf et al (2). Fluorescent lipid probes (5) were used that enabled the consistent resolution of two diffusion coefficients over a range of conditions.…”

supporting

confidence: 89%

“…The mobile fractions measured for both probes were 0.4-0.7. Similar results were observed in a study of lateral diffusion of plasma membrane components in protoplasts from suspensioncultured rose cells (4).…”

supporting

confidence: 88%

“…The present observations also shed light on the heterogeneous diffusion characteristics that have been observed for lipids in the plasma membranes of various protoplasts (2)(3)(4). In all of these previous studies, protoplasts were studied in standard media that have osmolalities around 700 mmol/kg or higher.…”

Section: Resultssupporting

confidence: 58%

“…When present at >33 mol %, cholesterol molecules are able to touch each other and are thought to form cholesterol-rich domains some tens of nanometers in size (23). Since sterols are more abundant in the plasma membrane than in endomembranes (24), it is likely that stressed protoplasts (Table 3) (3,4) and the survival of cells under freezing conditions (20,24).…”

Section: Resultsmentioning

confidence: 99%

“…Fluorescence photobleaching recovery experiments were performed as described (3)(4)(5), except for replacement of the argon-ion laser by a helium-cadmium laser operating at 441.6 nm in the present work. Appropriate bandpass filters and a dichroic beamsplitter for use with this laser line were designed and supplied by Omega Optical (Brattleboro, VT).…”

Section: (3)mentioning

confidence: 99%

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Selective osmotic effect on diffusion of plasma membrane lipids in maize protoplasts.

Furtula

Khan²,

Nothnagel³

1990

Proc. Natl. Acad. Sci. U.S.A.

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Osmotic levels in the range typically used during plant protoplast isolation and incubation were investigated with regard to effects on the lateral diffusion of lipid probes in the plasma membrane. The lateral diffusion coefficient of a fluorescent sterol probe in the plasma membrane of maize (Zea mays L.) root protoplasts in a medium containing 0.45 M mannitol was 4 times faster than when the medium contained 0.9 M mannitol. The lateral diffusion coefficient of a fluorescent phospholipid probe, however, did not change over this range of mannitol concentrations. Similar diffusion characteristics were observed when the medium contained trehalose instead of mannitol. Slower lateral diffusion of the sterol probe at higher osmolality was also observed when KCI/CaCl2-based osmotic media were used with protoplasts isolated by a mechanical, rather than by an enzymic, method. Extraction and quantitation of total lipids from protoplasts showed that both the phospholipid and sterol contents per protoplast decreased with increasing osmolality, while the sterol/phospholipid ratio increased. These results demonstrate that osmotic stress induces selective changes in both the composition and biophysical properties of plant membranes.Evidence for the existence of lipid domains in plant and other biological membranes has come from a variety of experiments (1). One source of such evidence has been studies of lipid dynamics involving long-range motions-i.e., motions over distances that are large compared to molecular dimensions. Use of fluorescence techniques to measure such lipid motions in the plasma membrane of plant protoplasts was first reported by Metcalf et al. (2). In that work, fluorescence redistribution after photobleaching was used to measure the lateral diffusion of various fluorescent lipid probes in protoplasts from suspension-cultured soybean cells. Two diffusion coefficients, one in the 2-5 x 1010 cm2/sec range and another in the 1-6 x 10-9 cm2/sec range, were simultaneously observed under certain conditions of temperature and probe concentration. The mobile fraction, summed for the two diffusing species, was in the 0.56-0.76 range-i.e., a significant portion of the lipid probe moved so slowly as to appear immobile on the time scale of the experiment. These heterogeneous diffusion parameters were interpreted as evidence for the existence of lipid domains in the plasma membrane of soybean protoplasts (1).Fluorescence photobleaching recovery experiments carried out in our laboratory (3, 4) have yielded results that are comparable to those reported by Metcalf et al. (2). Fluorescent lipid probes (5) were used that enabled the consistent resolution of two diffusion coefficients over a range of conditions. The lateral diffusion coefficient of a fluorescent sterol probe in the plasma membrane of root cortical protoplasts from two different maize lines was 3-7 x 10-10 cm2/sec for temperatures in the 3.50C-370C range (3). Over the same temperature range, the diffusion coefficient of a fluorescent phospholipid prob...

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supporting

confidence: 89%

supporting

confidence: 88%

Section: Resultssupporting

confidence: 58%

Section: Resultsmentioning

confidence: 99%

Section: (3)mentioning

confidence: 99%

See 3 more Smart Citations

Selective osmotic effect on diffusion of plasma membrane lipids in maize protoplasts.

Furtula

Khan²,

Nothnagel³

1990

Proc. Natl. Acad. Sci. U.S.A.

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Plant membrane biophysics in development and senescence

Leshem

1992

Plant Membranes

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Lipid lateral mobility in the plasma membrane of whole plant cells

Serpe

Nothnagel

1996

Pflugers Arch.

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Fluorescence photobleaching recovery was used to measure the apparent lateral diffusion coefficient and mobile fraction of a fluorescent lipid probe in the plasma membranes of whole plant cells, i.e., in the presence of cell walls. Interfering fluorescence from the cell wall was reduced by extensive washing and then subtracted from the recovery recordings. Mobility characteristics of plasma membrane lipids in whole cells were found to be very similar to those in protoplasts.

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

Cited by 16 publications

References 47 publications

Selective osmotic effect on diffusion of plasma membrane lipids in maize protoplasts.

Selective osmotic effect on diffusion of plasma membrane lipids in maize protoplasts.

Plant membrane biophysics in development and senescence

Lipid lateral mobility in the plasma membrane of whole plant cells

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