Incorporation of lipids into cellular membranes—A spin-label assay

Marsh, D.; Pellkofer, R.; Hoffmann-Bleihauer, P.; Sandhoff, Konrad

doi:10.1016/0003-2697(82)90272-x

Cited by 10 publications

(8 citation statements)

References 5 publications

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“…These two sites differ in their mobility; the probe in bulk sites is more mobile than that bound to the protein because of the mutual interaction through their hydrophobic segments. The focus of many works [see 17,19,20,26,27] has been the study of the lipidprotein interaction. In general, they have provided information about: i) the stoichiometry of the lipid -protein association, ii) the specificity of such association, iii) the conformational order in the lipid -protein interface and iv) the fatty acid exchange between general position and position bound to the protein.…”

Section: Figurementioning

confidence: 99%

Spin label electron paramagnetic resonance study in thylakoid membranes from a new herbicide-resistant D1 mutant from soybean cell cultures deficient in fatty acid desaturation

Yruela

Alfonso

García-Rubio

et al. 2001

Biochimica et Biophysica Acta (BBA) - Biomembranes

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The effect of fatty acid desaturation on lipid fluidity in thylakoid membranes isolated from the STR7 mutant was investigated by electron paramagnetic resonance (EPR) using spin label probes. The spectra of both 5- and 16-n-doxylstearic acid probes were measured as a function of the temperature between 10 and 305 K and compared to those of the wild type. This complete thermal evolution provides a wider picture of the dynamics. The spectra of the 5-n-doxylstearic acid probe as well as their temperature evolution were identical in both STR7 mutant and wild type thylakoids. However, differences were found with the 16-n-doxylstearic acid probe at temperatures between 230 and 305 K. The differences in the thermal evolution of the EPR spectra can be interpreted as a 5-10 K shift toward higher temperatures of the probe motional rates in the STR7 mutant as compared with that in the wild type. At temperatures below 230 K no differences were observed. The results indicated that the lipid motion in the outermost region of the thylakoids is the same in the STR7 mutant as in the wild type while the fluidity in the inner region of the STR7 mutant membrane decreases. Our data point out a picture of the STR7 thylakoid membrane in which the lipid motion is slower most probably as a consequence of fatty acid desaturation deficiency.

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Section: Figurementioning

confidence: 99%

Spin label electron paramagnetic resonance study in thylakoid membranes from a new herbicide-resistant D1 mutant from soybean cell cultures deficient in fatty acid desaturation

Yruela

Alfonso

García-Rubio

et al. 2001

Biochimica et Biophysica Acta (BBA) - Biomembranes

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“…4. These show a flexibility gradient between the three different label positions, which is characteristic of this type of label in a lipid bilayer environment (Marsh, 1981~). With all three labels there was no significant effect on the order parameter up to 60 pM melittin, whereas at higher concentrations the order parameter increased considerably with increasing melittin concentration.…”

Section: Esr Measurementsmentioning

confidence: 80%

“…Spin label order parameters were calculated from the outer and inner splittings in the ESR spectra by using the methods and corrections described by Marsh (1981a,b). The spectral lines were rather broad in some cases, indicating that the spectra may be approaching the slow motion region, in which case the apparent measured order parameters would be dependent on the rate of motion as well as the amplitude of motion (Marsh, 1981~).…”

Section: Esr Measurementsmentioning

confidence: 99%

“…The order parameter is a measure of the angular amplitude of motion of the lipid chain at the position of the spin label (Marsh, 1981~). The values obtained for the sphingomyelin spin labels incorporated into the SYM are given in Fig.…”

Section: Esr Measurementsmentioning

confidence: 99%

“…In order to estimate the amount of spin-labeled sphingomyelin incorporated into SYM, the spectrum obtained from vesicles prepared from pure 13-SPMSL was subtracted from the spectra obtained from 13-SPMSL incorporated into SYM. A detailed discussion of this method of assaying the degree of incorporation of spin-labeled lipids into membranes is found in Marsh et al (1981). Table 1 shows that increasing melittin concentrations caused increasing insertion of the sphingomyelin derivative into the membranes, reaching 90% in the presence of 100 p M melittin (second ESR spectra were obtained as described in Materials and Methods.…”

Section: Esr Measurementsmentioning

confidence: 99%

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Melittin Stimulates Incorporation and Degradation of Sphingomyelin in Synaptosomal Plasma Membranes

Pellkofer

Marsh

Hoffmann-Bleihauer

et al. 1982

Journal of Neurochemistry

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Melittin enhanced sphingomyelin (SPM) degradation by the neutral membrane-bound sphingomyelinase from calf brain synaptosomal plasma membranes (SYM) up to 20-fold. Melittin in concentrations as high as 100 microM did not significantly alter membrane fluidity of SYM as measured by fluorescence depolarization and electron spin resonance (ESR) using diphenylhexatriene and a doxyl derivative of SPM, respectively. In the concentration range 100--1000 microM, melittin was observed to rigidify SYM. The incorporation of SPM derivatives into the lipid bilayer of SYM was demonstrated by ESR measurements. Melittin enhanced the uptake of SMP-derivatives into SYM.

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Structural and Functional Modulation of Ion Channels by Specific Lipids: from Model Systems to Cell Membranes

Fernández

Poveda

Encinar

et al.

Springer Series in Biophysics

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Incorporation of lipids into cellular membranes—A spin-label assay

Cited by 10 publications

References 5 publications

Spin label electron paramagnetic resonance study in thylakoid membranes from a new herbicide-resistant D1 mutant from soybean cell cultures deficient in fatty acid desaturation

Spin label electron paramagnetic resonance study in thylakoid membranes from a new herbicide-resistant D1 mutant from soybean cell cultures deficient in fatty acid desaturation

Melittin Stimulates Incorporation and Degradation of Sphingomyelin in Synaptosomal Plasma Membranes

Structural and Functional Modulation of Ion Channels by Specific Lipids: from Model Systems to Cell Membranes

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