Solubility characteristics of Micrococcus lysodeikticus membrane components in detergents and chaotropic salts analyzed by immunoelectrophoresis.

In order to evalute the effectiveness and selectivity of various reagents in the solubilization of bacterial membranes, membranes of Micrococcus lysodeikticus were treated with detergents and chaotropic agents. The composition of the extracts so obtained was analyzed by rocket and two-dimensional immunoelectrophoretic techniques. Recoveery of succinate-, malate-, and reduced nicotinamide adenine dinucleotide- (NADH) dehydrogenases, ATPase, succinylated lipomannan and cytochromes in the extracts was measured. Treatment with a variety of non-denaturing detergents produced extracts that were generally qualitatively uniform although quantitative differences were observed. The degree of extraction of various components was correlated with the hydrophile-lipophile balance. Several chaotropic agents were also evaluated as reagents for membrane solubilization. These agents were less effective in extraction of bulk protein, but produced extracts enriched in some membrane components.

[1]  T. Steck,et al.  Selective solubilization of proteins and phospholipids from red blood cell membranes by nonionic detergents. , 1973, Journal of supramolecular structure.

[2]  P. Owen,et al.  Immunochemical analysis of inner and outer membranes of Escherichia coli by crossed immunoelectrophoresis , 1978, Journal of bacteriology.

[3]  A. Maddy.  The properties of the protein of the plasma membrane of ox erythrocytes. , 1966, Biochimica et biophysica acta.

[4]  Y. Anraku,et al.  Solubilization of a functionally active proline carrier from membranes of Escherichia coli with an organic solvent. , 1977, Biochemical and biophysical research communications.

[5]  S. Hjertén,et al.  Some observations on the choice of detergent for solubilization of the human erythrocyte membrane. , 1977, The Biochemical journal.

[6]  R. Smith,et al.  Lowry determination of protein in the presence of Triton X-100. , 1975, Analytical biochemistry.

[7]  M. Schor,et al.  Release and purification of Micrococcus lysodeikticus ATPase from membranes extracted with n-butanol. , 1974, Biochimica et biophysica acta.

[8]  J. Freer,et al.  Membrane-associated ATPase activity from Micrococcus lysodeikticus. , 1968, Biochimica et biophysica acta.

[9]  J. B. Finean,et al.  Detergent extraction of erythrocyte ghosts. Comparison of residues after cholate and Triton X-100 treatments. , 1976, Biochimica et biophysica acta.

[10]  J. Donovan,et al.  Batch purification of ovomucoid and characterization of the purified product. , 1971, Biochemistry.

[11]  O. Bjerrum,et al.  The immunochemical approach to the characterization of membrane proteins. Human erythrocyte membrane proteins analysed as a model system. , 1976, Biochimica et biophysica acta.

[12]  F. L. Crane,et al.  Fractionation of mitochondrial membranes with sodium deoxycholate. , 1972, Biochimica et biophysica acta.

[13]  D. Maclennan.  Purification and properties of an adenosine triphosphatase from sarcoplasmic reticulum. , 1970, The Journal of biological chemistry.

[14]  S. Hjertén,et al.  Purification and characterization of spiralin, the main protein of the Spiroplasma citri membrane. , 1977, Biochimica et biophysica acta.

[15]  M. Salton,et al.  Immunological Properties of Micrococcus lysodeikticus Membranes , 1971, Journal of bacteriology.

[16]  N. Harboe,et al.  23. Immunization, Isolation of Immunoglobulins, Estimation of Antibody Titre , 1973, Scandinavian journal of immunology. Supplement.

[17]  A. Helenius,et al.  Solubilization of membranes by detergents. , 1975, Biochimica et biophysica acta.

[18]  B. Litman,et al.  Effect of alterations in the amphipathic microenvironment on the conformational stability of bovine opsin. 1. Mechanism of solubilization of disk membranes by the nonionic detergent, octyl glucoside. , 1978, Biochemistry.

[19]  T. Flatmark,et al.  Effect of the hydrophile-lipophile balance of non-ionic detergents (Triton X-series) on the solubilization of biological membranes and their integral b-type cytochromes. , 1976, Biochimica et biophysica acta.

[20]  C. Urban,et al.  Selective extraction of gonococcal envelope antigens with thiocyanates , 1978 .

[21]  S. Hjertén,et al.  Localization of the Tween 20-soluble membrane proteins of Acholeplasma laidlawii by crossed immunoelectrophoresis. , 1974, Journal of molecular biology.

[22]  J. Oppenheim,et al.  The use of lectins in the quantitation and analysis of macromolecules by affinoelectrophoresis. , 1977, Analytical biochemistry.

[23]  P. Owen,et al.  Membrane Asymmetry and Expression of Cell Surface Antigens of Micrococcus lysodeikticus Established by Crossed Immunoelectrophoresis , 1977, Journal of bacteriology.

[24]  S. Hjertén,et al.  Selective solubilization with Tween 20 of membrane proteins from Acholeplasma laidlawaii. , 1972, Biochimica et biophysica acta.

[25]  J. Hays,et al.  Irreversibel inactivation of the membrane-bound enzyme IIlac of the lactose phosphotransferase system of Staphylococcus aureus by triton X-100 and protection by substrates. , 1977, Biochimica et biophysica acta.

[26]  R. Niederman,et al.  Membranes of Rhodopseudomonas sphaeroides. IV. Assembly of chromatophores in low-aeration cell suspensions. , 1976, Biochimica et biophysica acta.

[27]  R. Egan.  Hydrophile-lipophile balance and critical micelle concentration as key factors influencing surfactant disruption of mitochondrial membranes. , 1976, The Journal of biological chemistry.

[28]  D. Miller,et al.  Total solubilization of erythrocyte membranes bo nonionic detergents. , 1970, Biochemical and biophysical research communications.

[29]  C. Smyth,et al.  Antigenic analysis of Neisseria gonorrhoeae by crossed immunoelectrophoresis , 1976, Infection and immunity.

[30]  S. Razin,et al.  Solubilization of mycoplasma membranes by the nonionic detergent Triton X‐100 , 1969, FEBS letters.

[31]  J. Freer,et al.  Electron transport components localized in a lipid-depleted sheet isolated from Micrococcus lysodeikticus membranes by deoxycholate extraction. , 1968, Biochemical and biophysical research communications.

[32]  M. Salton,et al.  Affinity chromatography of succinate dehydrogenase from the membranes of Micrococcus lysodeikticus. , 1975, Preparative biochemistry.

[33]  B. Kê,et al.  A photochemically active particle derived from chloroplasts by the action of the detergent Triton X-100. , 1966, The Journal of biological chemistry.

[34]  W. Hengstenberg,et al.  Morphological changes in staphylococcal cytoplasmic membrane due to action of non-ionic detergent Triton X-100. , 1973, Experimental cell research.

[35]  J. Freer,et al.  Composition of the membranes isolated from several Gram-positive bacteria. , 1965, Biochimica et biophysica acta.

[36]  O. Bjerrum,et al.  Crossed immunoelectrophoresis of human erythrocyte membrane proteins. Immunoprecipitation patterns for fresh and stored samples of membranes extensively solubilized with non-ionic detergents. , 1974, Biochimica et biophysica acta.

[37]  J. Hays,et al.  Selective, reversible inhibition of the lactose phosphotransferase system of Staphylococcus aureus by dodecyl sulfate and deoxycholate. , 1977, Archives of biochemistry and biophysics.

[38]  M. Collins,et al.  Membranes of Rhodospirillum rubrum: isolation and physicochemical properties of membranes from aerobically grown cells , 1976, Journal of bacteriology.

[39]  J. Rydström,et al.  Selective solubilization of the components of the mitochondrial inner membrane by lysolecithin. , 1976, Biochimica et biophysica acta.

[40]  N. R. Bergquist.  The Pulsed Dye Laser as a Light Source for the Fluorescent Antibody Technique , 1973, Scandinavian journal of immunology.

[41]  K. A. Davis,et al.  Purification, molecular properties, and amino acid composition of the subunits of Rhodospirillum rubrum succinate dehydrogenase. , 1977, Archives of biochemistry and biophysics.

[42]  M. Salton,et al.  Characteristics of a lipid-rich NADH dehydrogenase-containing particulate fraction obtained from Micrococcus lysodeikticus membranes. , 1970, Biochimica et biophysica acta.

[43]  E. Munn,et al.  A simple and rapid method for the preparation of adenosine triphosphatase from submitochondrial particles. , 1975, The Biochemical journal.

[44]  K. A. Davis,et al.  Succinate dehydrogenase. I. Purification, molecular properties, and substructure. , 1971, Biochemistry.

[45]  C. Laurell,et al.  ANTIGEN-ANTIBODY CROSSED ELECTROPHORESIS. , 1965, Analytical biochemistry.

[46]  P. Owen,et al.  Submicrogram quantitation of an acidic polysaccharide by rocket immunoelectrophoresis and rocket affinoelectrophoresis. , 1976, Analytical biochemistry.

[47]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[48]  Henri Wróblewski.  Dissolution sélective de protéines de la membrane de Spiroplasma citri par le désoxycholate de sodium , 1975 .

[49]  R. Ernst,et al.  Solubilization of membrane proteins by sulfobetaines, novel zwitterionic surfactants. , 1978, Analytical biochemistry.

[50]  J. Strominger,et al.  D-alanine carboxypeptidase from Bacillus subtilis membranes. I. Purification and characterization. , 1973, The Journal of biological chemistry.

[51]  H. Crone.  THE DISSOCIATION OF RAT BRAIN MEMBRANES BEARING ACETYLCHOLINESTERASE BY THE NON‐IONIC DETERGENT TRITON X‐100 AND AN EXAMINATION OF THE PRODUCT , 1971, Journal of neurochemistry.

[52]  A. Martonosi.  Sarcoplasmic reticulum. IV. Solubilization of microsomal adenosine triphosphatase. , 1968, The Journal of biological chemistry.