The properties of the protein of the plasma membrane of ox erythrocytes

Maddy, A.H.

doi:10.1016/0304-4165(66)90166-8

Cited by 193 publications

(52 citation statements)

References 12 publications

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“…Proteins were extracted from isolated plasma membranes using n-butanol according to the method of Maddy (1966). Membranes (10 mg/ml protein) in hypotonic buffer containing 0.2 mMMgC12 were mixed with 0.75 vol.…”

Section: Methodsmentioning

confidence: 99%

Characterization of Membrane Components of the Erythrocyte Involved in Vesicular Stomatitis Virus Attachment and Fusion at Acidic pH

Mastromarino

Conti

Goldoni

et al. 1987

Journal of General Virology

130

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SUMMARYGoose erythrocyte membranes were isolated and tested for their ability to compete with red cell receptors for vesicular stomatitis virus (VSV) attachment and fusion at acidic pH. Crude membranes, solubilized with Triton X-100, Tween 80 and octyl-/~-Dglucopyranoside, showed a dose-dependent inhibitory effect on virus binding and haemolysis. The chemical nature of the active molecules was investigated by enzyme digestion and by separation of purified components. Only the lipid moiety, specifically phospholipid and glycolipid, was found to inhibit VSV attachment; a more detailed analysis of these molecules showed that phosphatidylinositol, phosphatidylserine and GM3 ganglioside were responsible for the inhibitory activity and could therefore represent VSV binding sites on goose erythrocyte membranes. Removal of negatively charged groups from these molecules by enzymic treatment significantly reduced their activity, suggesting that electrostatic interactions play an important role in the binding of VSV to the cell surface. Enzymic digestion of whole erythrocytes confirmed the involvement of membrane lipid molecules in the cell surface receptor for VSV.

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Section: Methodsmentioning

confidence: 99%

Characterization of Membrane Components of the Erythrocyte Involved in Vesicular Stomatitis Virus Attachment and Fusion at Acidic pH

Mastromarino

Conti

Goldoni

et al. 1987

Journal of General Virology

130

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“…It meant that the solubilizing solutions (b, d and e) were superior to the others. A number of attempts have been adopted for solubilization of red cell membrane; organic solvents (Maddy 1966;Hamaguchi and Cleve 1972), non-ionic detergents (Bjerrum and Lundahl 1974), ionic detergents (Corry and Stone 1969). Of these reagents, ionic detergent such as SDS could extract most of the proteins on the membrane, whereas integral proteins of the membrane were difficult to isolate by the other extractions.…”

Section: Discussionmentioning

confidence: 99%

Immunoelectrophoretic analyses of antigenic and human specific proteins of red cell membrane.

Iwasa

Yokoi

Kanemitsu

et al. 1982

Tohoku J. Exp. Med.

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Rabbit antisera were raised for red cell stroma extracted with each of the following solutions; 5 mM phosphate buffer pH 8.0 (A antigen), 5 mM EDTA containing 5 mM 2-mercaptoethanol (B), 1/15 M phosphate buffered saline (C), 0.05% SDS (D), 50 mM Tris-HC1 buffer pH 8.0 containing 1% SDS, 1 mM EDTA and 1% 2-mercaptoethanol (E) and 1% Triton X-100 (F). Crossed immunoelectrophoresis showed that E antigen produced relatively numerous precipitation lines, 7-9, against each of the antisera, suggesting that E solution was superior to the others for membrane solubilization. However, the Ouchterlony test demonstrated potent precipitin activity in anti-C, and the antiserum absorbed with monkey stroma which was extracted with F solution was proved to have strict human specificity. By adopting the antiserum to affinity chromatography, human specific antigens were isolated. The antigens gave four bands moving faster than Band 5 at SDS-polyacrylamide gel electrophoresis none of which was stained by Schiff's reagent. Using absorbed anti-C, human specificity of blood stains kept up to for two years could be identified. red cell membrane; human specific antigen; electrophoresis; membrane solubilization In forensic practice, species-identification of blood stains found at the scene was performed by immunoassay with anti-human hemoglobin. Recently, human specificity of each of plasma proteins was investigated by immunochemical methods (James 1965;Esteves and Binaghi 1972;Bauer 1974). On the other hand, little information about the antigens on or in red cell membrane has been presented. It was well known that the animals immunized with red cells of different species produced heteroagglutinins of various titers, indicating that species specific antigens were contained in red cell membrane.Recent advances in solubilizing red cell membrane revealed some constituents with antigenic activity of blood group or of enzymes (Madly and Dunn 1976). Bjerrum and Lundahl (1974) and Bjerrum et al. (1975) analyzed antigenicity of constituents of human red cells by two dimensional electrophoresis in which 19 membrane-specific immunoprecipitates were demonstrated.In the present study, we compare the solubilizing ability of various solutions

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“…However, since their antigenicity and absorption capacity was comparable to that of untreated membranes, this probably reflects the failure of the detergent to lyse the butanol-treated membranes rather than a loss of antigens. The butanol-extracted lipid neither reacted in gel diffusion tests nor absorbed MI or IHA antibody from antiserum to whole organisms ( Maddy (1966) or Zwaal & van Deenen (1968) or with lithium salts contained only small amounts of protein and gave, at best, weak reactions in gel diffusion tests with antiserum to whole organisms. However, urea and phenolic extracts gave lines corresponding to components 1 and 3.…”

Section: Membrane Antigen8 Of Mycoplasma Hominismentioning

confidence: 90%

“…The insoluble pellet and interfacial material was resuspended in the aqueous layer, re-extracted with two volumes of butanol and centrifuged as before. Maddy (1966) and Zwaal & van Deenen (1968) for preparing soluble protein or lipoprotein extracts from mammalian erythrocyte ghosts were used with M. hominis membrane suspensions. Treatment with lithium chloride or lithium bromide at concentrations from 0 1 to 8 M was also tried.…”

Section: Serological Testsmentioning

confidence: 99%

Membrane antigens ofMycoplasma hominis

Hollingdale

Lemcke

1972

J. Hyg.

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SUMMARYExtraction of membranes of Mycoplasma hominis with n-butanol showed that antigenicity was associated with the non-lipid residue, which probably consisted mainly of protein, and not with the lipid itself.Since many membrane proteins are hydrophobic, membranes were rendered soluble in various ways. Extraction with urea or phenol was the most successful, yielding extracts which were both antigenic and serologically reactive. The urea extract could not be fractionated by polyacrylamide disk electrophoresis or by column chromatography. However, serologically active components identified by gel diffusion were separated from detergent-lysed membranes by polyacrylamide disk electrophoresis. The activities of antisera against these fractions suggested that indirect-haemagglutinating or metabolic-inhibiting antibodies can be directed against several different membrane antigens. However, the antigens identified by gel diffusion probably do not represent all the components participating in indirect haemagglutination.Treatment of membrane suspensions with heat, alkali, periodate and various enzymes showed that the four components identified by gel diffusion could be distinguished by their differing stabilities and properties. On the basis of their lability and susceptibility to proteolytic enzymes, two were identified as proteins.

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The properties of the protein of the plasma membrane of ox erythrocytes

Cited by 193 publications

References 12 publications

Characterization of Membrane Components of the Erythrocyte Involved in Vesicular Stomatitis Virus Attachment and Fusion at Acidic pH

Characterization of Membrane Components of the Erythrocyte Involved in Vesicular Stomatitis Virus Attachment and Fusion at Acidic pH

Immunoelectrophoretic analyses of antigenic and human specific proteins of red cell membrane.

Membrane antigens ofMycoplasma hominis

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