The characterisation of membrane proteins by centrifugation and gel electrophoresis a comparison of proteins prepared by different methods

Maddy, A.H.; Dünn, Michael J.; Kelly, Padraig

doi:10.1016/0005-2736(72)90247-7

Cited by 40 publications

(11 citation statements)

References 31 publications

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“…Small unilamellar liposomes (SUV) were composed of lipids extracted from erythrocyte membranes (RBCL) according to the method described by Maddy et al (1972), dissolved in a chloroform:methanol solvent and of DPPC. All lipids were evaporated to dryness under nitrogen.…”

Section: Methodsmentioning

confidence: 99%

Biophysical Mechanism of the Protective Effect of Blue Honeysuckle (Lonicera caerulea L. var. kamtschatica Sevast.) Polyphenols Extracts Against Lipid Peroxidation of Erythrocyte and Lipid Membranes

et al. 2014

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The aim of the present research was to determine the effect of blue honeysuckle fruit and leaf extracts components on the physical properties of erythrocyte and lipid membranes and assess their antioxidant properties. The HPLC analysis showed that the extracts are rich in polyphenol anthocyanins in fruits and flavonoids in leaves. The results indicate that both extracts have antioxidant activity and protect the red blood cell membrane against oxidation induced by UVC irradiation and AAPH. The extracts do not induce hemolysis and slightly increase osmotic resistance of erythrocytes. The research showed that extracts components are incorporated mainly in the external part of the erythrocyte membrane, inducing the formation of echinocytes. The values of generalized polarization and fluorescence anisotropy indicate that the extracts polyphenols alter the packing arrangement of the hydrophilic part of the erythrocyte and lipid membranes, without changing the fluidity of the hydrophobic part. The DSC results also show that the extract components do not change the main phase transition temperature of DPPC membrane. Studies of electric parameters of membranes modified by the extracts showed that they slightly stabilize lipid membranes and do not reduce their specific resistance or capacity. Examination of IR spectra indicates small changes in the degree of hydration in the hydrophilic region of liposomes under the action of the extracts. The location of polyphenolic compounds in the hydrophilic part of the membrane seems to constitute a protective shield of the cell against other substances, the reactive forms of oxygen in particular.

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Section: Methodsmentioning

confidence: 99%

Biophysical Mechanism of the Protective Effect of Blue Honeysuckle (Lonicera caerulea L. var. kamtschatica Sevast.) Polyphenols Extracts Against Lipid Peroxidation of Erythrocyte and Lipid Membranes

et al. 2014

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“…Fresh blood was taken each time to a physiological solution of sodium chloride with heparin added. The lipids were extracted from erythrocyte membranes according to the method described by Maddy et al [23]. Erythrocyte ghosts were obtained according to Dodge et al [24].…”

Section: Methodsmentioning

confidence: 99%

Interaction of selected anthocyanins with erythrocytes and liposome membranes

Bonarska-Kujawa

Pruchnik

Kleszczyńska

2012

Cellular and Molecular Biology Letters

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Anthocyanins are one of the main flavonoid groups. They are responsible for, e.g., the color of plants and have antioxidant features and a wide spectrum of medical activity. The subject of the study was the following compounds that belong to the anthocyanins and which can be found, e.g., in strawberries and chokeberries: callistephin chloride (pelargonidin-3-O-glucoside chloride) and ideain chloride (cyanidin-3-O-galactoside chloride). The aim of the study was to determine the compounds' antioxidant activity towards the erythrocyte membrane and changes incurred by the tested anthocyanins in the lipid phase of the erythrocyte membrane, in liposomes composed of erythrocyte lipids and in DPPC, DPPC/cholesterol and egg lecithin liposomes. In particular, we studied the effect of the two selected anthocyanins on red blood cell morphology, on packing order in the lipid hydrophilic phase, on fluidity of the hydrophobic phase, as well as on the temperature of phase transition in DPPC and DPPC/cholesterol liposomes. Fluorimetry with the Laurdan and Prodan probes indicated increased packing density in the hydrophilic phase of the membrane in the presence of anthocyanins. Using the fluorescence probes DPH and TMA-DPH, no effect was noted inside the hydrophobic phase of the membrane, as the lipid bilayer fluidity was not modified. The compounds slightly lowered the phase transition temperature of phosphatidylcholine liposomes. The study has shown that both anthocyanins are incorporated into the outer region of the erythrocyte membrane, affecting its shape and lipid packing order, which is reflected in the increasing number of echinocytes. The investigation proved that the compounds penetrate only the outer part of the external lipid layer of liposomes composed of erythrocyte lipids, DPPC, DPPC/cholesterol and egg lecithin lipids, changing its packing order. Fluorimetry studies with DPH-PA proved that the tested anthocyanins are very effective antioxidants. The antioxidant activity of the compounds was comparable with the activity of Trolox ® .

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“…Membranes were prepared by the method of Shimomura et al (4). After butanol extraction (18) and dialysis against 10 mM Tris/1 mM EDTA, pH 7.0 proteins were equilibrated with 40 ml of QA Trisacryl (Pharmacia) in the same buffer for 4 hr. The resin was then poured into a column, washed in buffer, and eluted with 0.3 M NaCl in the same buffer; the proteins were then concentrated by ultrafiltration (Centriprep 3; Amicon).…”

Section: Materals and Methodsmentioning

confidence: 99%

Antibodies to a peptide from the maize auxin-binding protein have auxin agonist activity.

Venis

Napier

Barbier‐Brygoo

et al. 1992

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

123

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The major auxin-binding protein in maize membranes is thought to function as a physiological receptor. From earlier information, including the use of site-directed irreversible inhibitors, several of the amino acids likely to form part of the active auxin-binding site were provisionally assigned. Inspection of the amino acid sequence of the auxinbini protein showed a short region containing all but one of these ano acids. We find that antisera raised against a synthetic peptide encompassing this region recognize all isoforms of the mize auxin-binding protein together with homologous polypeptides in other species. We further find that the antibodies hyperpolarIze protoplast transmembrane potential in an auxin-like manner. We conclude that these antibodies display auxin agonist activity and that we have identified an essential portion of the auxin-binding site. 7208The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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The characterisation of membrane proteins by centrifugation and gel electrophoresis a comparison of proteins prepared by different methods

Cited by 40 publications

References 31 publications

Biophysical Mechanism of the Protective Effect of Blue Honeysuckle (Lonicera caerulea L. var. kamtschatica Sevast.) Polyphenols Extracts Against Lipid Peroxidation of Erythrocyte and Lipid Membranes

Biophysical Mechanism of the Protective Effect of Blue Honeysuckle (Lonicera caerulea L. var. kamtschatica Sevast.) Polyphenols Extracts Against Lipid Peroxidation of Erythrocyte and Lipid Membranes

Interaction of selected anthocyanins with erythrocytes and liposome membranes

Antibodies to a peptide from the maize auxin-binding protein have auxin agonist activity.

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