Isoelectric Focusing of Microbial Proteins

  • William W. Yotis
Part of the Biological Separations book series (BIOSEP)


The application of isoelectric focusing for the isolation and characterization of microbial proteins is increasing slowly. While such methods as electrophoresis, chromatography, ultracentrifugation, as well as fractionation of microbial proteins by salts and organic solvents are all occupying a prominent place in analytical biochemistry, today isoelectric focusing ranks as one of the leading tools in the analytical laboratory. The real strength of isoelectric focusing lies in its ability to provide separation, quantitation, and characterization of quite closely related amphoteric compounds. Proteins varying by as little as 0.02 pH units at their isoelectric point have been clearly resolved.


Isoelectric Point Serratia Marcescens Sucrose Density Gradient Staphylococcal Enterotoxin Rous Sarcoma Virus 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Ahlgren, E., and Eriksson, K.-E. 1967, Characterization of cellulases and related enzymes by isoelectric focusing, gel filtration and zone electrophoresis. II. Studies on Stereum sanguinolentum, Fomes annosus and Chrysosporium lignorum enzymes, Acta Chem. Scand. 21:1193.CrossRefGoogle Scholar
  2. Ahlgren, E., Eriksson, K.-E., and Vesterberg, O. 1967, Characterization of cellulases and related enzymes by isoelectric focusing, gel filtration and zone electrophoresis. I. Studies on Aspergillus enzymes, Acta Chem. Scand. 21:937.CrossRefGoogle Scholar
  3. Ambler, J., Miller, J. N., and Orr, T. S. C. 1973, Characterization of an allergen extracted from Ascaris suum. Determination of the molecular weight, isoelectric point, amino acid composition and carbohydrate content of the native allergen, Immunochemistry 10:815.PubMedCrossRefGoogle Scholar
  4. Andersson, W. B., Schneider, A. B., Emmer, M., Perlman, R. L., and Pastan, I. 1971, Purification of and properties of the cyclic adenosine 3', 5'-monophosphate receptor protein which mediates cyclic adenosine 3', 5'-monophosphate-dependent gene transcription in Escherichia coli, J. Biol. Chem. 246:5929.Google Scholar
  5. Arbuthnott, J. P., and Beeley, J. A., 1975, “Isoelectric Focusing,“ Butterworths, London.Google Scholar
  6. Arens, A., Rauenbusch, E., Irion, E., Wagner, O., Bauer, K., and Kaufmann, W. 1970, Isolation and properties of l-asparaginases from Escherichia coli, Hoppe-Seyler’s Z. Physiol. Chem. 351:197.PubMedCrossRefGoogle Scholar
  7. Arvidson, S., Holme, T., and Lindholm, B. 1973, Studies on extracellular proteolytic enzymes from Staphylococcus aureus. I. Purification and characterization of one neutral and one alkaline protease, Biochim. Biophys. Acta 302:135.PubMedGoogle Scholar
  8. Augier, J., and Augier-Gibory, S. 1969, Analysis by electrofocusing of protein fractions present in the purified protein derivatives (PPD), Ann. Instit. Pasteur 117:768.Google Scholar
  9. Avadhani, N. G., and Buetow, D. E. 1972, Studies on messenger RNA from Euglena gracilis. I. Techniques of saline fractionation and characterization of messenger RNAs from dividing cells, Biochim. Biophys. Acta 262:488.PubMedGoogle Scholar
  10. Awdeh, Z. L., Williamson, A. R., and Askonas, B. A. 1968, Isoelectric focusing in polyacrylamide gel and its application to immunoglobuhns, Nature (Lond.) 219:66.CrossRefGoogle Scholar
  11. Bancroft, J. B., Hiebert, E., Rees, M. W., and Markham, R. 1968, Properties of cowpea chlorotic mottle virus, its protein and nucleic acid, Virology 34:224.PubMedCrossRefGoogle Scholar
  12. Barret, T., and Gould, H. J. 1973, Tissue and species specificity of non-histone chromatin proteins, Biochim. Biophys. Acta 294:165.Google Scholar
  13. Beers, W. H., and Reich, E. 1969, Isolation and characterization of Clostridium botulinum Type B toxin, J. Biol. Chem. 244:4473.PubMedGoogle Scholar
  14. Bernheimer, A., W., Auigad, L. S., and Shin, K. 1974, Staphylococcal sphingomyelinase (1-hemolysin), in “Advances in Staphylococcal Research” (W. W. Yotis, ed.), Ann. N. Y. Acad. Sci. 236:292.Google Scholar
  15. Birken, S., and Pisano, M. A. 1976, Purification and properties of a polyol dehydrogenase from Cephalosporium chrysogenum, J. Bacteriol. 125:225.PubMedGoogle Scholar
  16. Bischofberger, H., Hess, B., Röschlau, Pl., Wieker, H-J., and Zimmermann-Telschow, H. 1970, Amino acid composition and subunit structure of yeast pyruvate kinase, Hoppe-Seyler’s Z. Physiol. Chem. 351:401.PubMedCrossRefGoogle Scholar
  17. Boethling, R. S. 1975, Purification and properties of a serine protease from Pseudomonas maltophilia, J. Bacteriol. 121:933.PubMedGoogle Scholar
  18. Bosman, H. B. 1973, Protein catabolism. II. Identification of neutral and acidic proteolytic enzymes in Aspergillus niger, Biochim. Biophys. Acta 293:476.Google Scholar
  19. Brown, D. G., and Abrams, A. 1970, The proteins in membrane and cytoplasmic ribosomes of Streptococcus faecalis. An extra protein in the 50-S subparticles of cytoplasmic ribosomes, Biochim. Biophys. Acta 200:522.PubMedGoogle Scholar
  20. Brown, D. G., Baron, C., and Abrams, A. 1968, The separation of the ribosomal proteins of Streptococcus fecalis by isoelectric focusing. Amino acid composition of the total ribosomal proteins and of an acidic fraction, Biochim. Biophys. Acta 168:386.PubMedGoogle Scholar
  21. Campbell, H. A., and Mashburn, L. T. 1969, l-asparaginase EC-2 from Escherichia coli. Some substrate specificity characteristics, Biochemistry 8:3768.PubMedCrossRefGoogle Scholar
  22. Campbell, W. H., Orne-Johnson, W. H., and Burris, R. H. 1973, A comparison of the physical and chemical properties of four cytochromes c from Azotobacter vinelandii, Biochim. Biophys. Acta 135:617.Google Scholar
  23. Carlsson, J. 1970, A levansucrase from Streptococcus mutans, Caries Res. 4:97.PubMedCrossRefGoogle Scholar
  24. Carlsson, J., Newbrun, E., and Krasse, B. 1969, Purification and properties of dextransucrase from Streptococcus sanguis, Arch. Oral. Biol. 14:469.PubMedCrossRefGoogle Scholar
  25. Catsimpoolas, N. 1969, Immunoelectrofocusing in agarose gels, Clin. Chim. Acta 23:237.PubMedCrossRefGoogle Scholar
  26. Catsimpoolas, N. 1970, Isoelectrc focusing of proteins in gel media, Sep. Sci. 5:523.CrossRefGoogle Scholar
  27. Catsimpoolas, N. 1971, Analytical scanning isoelectrofocusing: Design and operation of an in situ scanning apparatus, Anal. Biochem. 44:411.PubMedCrossRefGoogle Scholar
  28. Catsimpoolas, N. 1973a, Isoelectrc focusing and isotachophoresis of proteins, Sep. Sci. 8:71.CrossRefGoogle Scholar
  29. Catsimpoolas, N. 1973b, Isoelectric Focusing and Isotachophoresis, Ann. N. Y. Acad. Sci. 209:1.Google Scholar
  30. Catsimpoolas, N. 1973c, Immunoelectrofocusing, Ann. N.Y. Acad. Sci. 209:144.CrossRefGoogle Scholar
  31. Catsimpoolas, N. 1973d, Transient state isoelectric focusing, in “Isoeiectric Focusing” (J. P. Arbuthnott and J. A. Beeley, eds.), pp. 58–73, Butterworths, London.Google Scholar
  32. Catsimpoolas, N. 1975, Isoelectrc focusing: Fundamental aspects, Sep. Sci. 10:55.CrossRefGoogle Scholar
  33. Catsimpoolas, N., Yotis, W. W., Griffith, A. L., and Rodbard, D. 1974, Transient state isoelectric focusing: Effect of zone load and carrier ampholyte concentration on the kinetics of defocusing and refocusing in sucrose density gradients, Arch. Biochem. Biophys. 163:113.PubMedCrossRefGoogle Scholar
  34. Chaiet, L., Kempf, A. J., Harman, R., Kaczka, E., Weston, R., Nollstadt, K., and Wolf, F. J. 1970, Isolation of a pure dextranase from Penicillium funiculosum, Appl. Microbiol. 20:421.PubMedGoogle Scholar
  35. Chang, P. C., and Dickie, N. 1971, Fractionation of staphylococcal enterotoxin B by isoelectric focusing, Biochim. Biophys. Acta 236:367.PubMedGoogle Scholar
  36. Chang, P. C., Yano, Y., Dighton, M., and Dickie, N. 1971, Fractionation of staphylococcal enterotoxin C2 by isoelectric focusing, Canad. J. Microbiol. 17:1367.CrossRefGoogle Scholar
  37. Cheng, A., Kwapinski, J. B. G., and Ronald, A. R. 1974, Isolation and immunological characterization of a unique toxic cytoplasmic polymer of type 1 Neisseria gonorrheae, Canad. J. Microbiol. 20:1163.CrossRefGoogle Scholar
  38. Chila, R., Doering, K. M., Domagk, G. F., and Rippa, M. 1973, A simplified procedure for the isolation for a highly active crystalline glucose-6-phosphate dehydrogenase from Candida utilis, Arch. Biochem. Biophys. 159:235.CrossRefGoogle Scholar
  39. Clark-Walker, G. D., and Lascelles, J. 1970, Cytochrome C550 from Spirillum itersonii: Purification and some properties, Arch. Biochem. Biophys. 136:153.PubMedCrossRefGoogle Scholar
  40. Colonna, W. J., Cano, F. R., and Lampen, O. J. 1975, Microheterogeneity of yeast invertase, Biochim. Biophys. Acta 236:293.Google Scholar
  41. Csopak, H., Jonsson, M., and Hallberg, B. 1972, Isoelectric fractionation of the Escherichia coli alkaline phosphatase and resolution of the purified enzyme into isoenzymes, Acta Chem. Scand. 26:2412.PubMedCrossRefGoogle Scholar
  42. Cunningham, M., and Beachey, E. H. 1975, Immunochemical properties of streptococcal M protein purified by isoelectric focusing, J. Immunol. 115:1002.PubMedGoogle Scholar
  43. Dale, G., and Latner, A. L. 1969, Isoelectric focusing of serum proteins in acrylamide gels followed by electrophoresis, Clin. Chim. Acta 24:61.PubMedCrossRefGoogle Scholar
  44. DeGraaf, F. K., Goedvolk-DeGroot, L. E., and Stouthamer, A. H. 1970, Purification of a bacteriocin produced by Enterobacter cloacae DF 13, Biochim. Biophys. Acta 221:566.Google Scholar
  45. Delisle, G., and Milazzo, F. H. 1970, The isolation of arylsulphatase isoenzymes from Pseudomonas aeruginosa, Biochim. Biophys. Acta 212:505.PubMedGoogle Scholar
  46. Dickie, N., Yano, Y., Robern, H., and Stauric, S. 1972, On the heterogeneity of staphylococcal enterotoxin C2, Canad. J. Microbiol. 18:801.CrossRefGoogle Scholar
  47. Domagk, G. F., Doering, M. K., and Chilla, R. 1973, Purification and properties of ribosephosphate isomerase from Candida utilis, Eur. J. Biochem. 38:259.PubMedCrossRefGoogle Scholar
  48. Eady, R. R., Smith, B. E., Cook, K. A., and Postgate, J. R. 1972, Nitrogenase of Klebsiella pneumoniae. Purification and properties of the component proteins, Biochem. J. 128:655.PubMedGoogle Scholar
  49. Ehrman, M., Cedar, H., and Schwartz, J. H. 1971, l-Asparaginase II of Escherichia coli. Studies on the enzymatic mechanism of action, J. Biol. Chem. 246:88.PubMedGoogle Scholar
  50. Epstein, I., and Grossowicz, N. 1975, Purification and properties of glutamate dehydrogenase from a thermophilic bacillus, J. Bacteriol. 122:1257.PubMedGoogle Scholar
  51. Esser, K., and Minuth, W. 1970, The phenoloxidases of the ascomycete Podospora anserina, Communication VI. Genetic regulation of the formation of laccase, Genetics 64:441.PubMedGoogle Scholar
  52. Falcoz-Kelly, F., Janin, J., Saari, J. C., Veron, M., Truffa-Bachi, P., and Cohen, G. N. 1972, Revised structure of aspartokinase I—homoserine dehydrogenase I of Escherichia coli K12. Evidence for four identical subunits, Eur. J. Biochem. 28:507.PubMedCrossRefGoogle Scholar
  53. Fawcett, J. S. 1973, Continuous-flow isoelectric focusing and isotachophoresis, Ann. N.Y. Acad. Sci. 209:112.PubMedCrossRefGoogle Scholar
  54. Feichter, A., Mian, F. A., Ris, H., and Halvorson, H. O. 1972, Characterization of insoluble protein fractions of mitochondria from Saccharomyces cerevisiae, J. Bacteriol. 109:855.Google Scholar
  55. Finkelstein, R. A., and LoSpalluto, J. J. 1970, Production of highly purified choleragen and choleragenoid, J. Infect. Dis. 121(Suppl.): S63.CrossRefGoogle Scholar
  56. Finlayson, G. R., and Chrambach, A. 1971, Isoelectric focusing in polyacrylamide gel and its preparative application, Anal. Biochem. 40:292.PubMedCrossRefGoogle Scholar
  57. Fodge, D. W., Gracy, R. W., and Harris, B. G. 1972, Studies on enzymes from parasitic helminths. I. Purification and physical properties of malic enzyme from the muscle tissue of Ascaris suum, Biochem. Biophys. Acta 268:271.PubMedGoogle Scholar
  58. Frere, J.-M., Ghuysen, J.-M., Perkins, H. R., and Nieto, M. 1973, Molecular weight and amino acid composition of the extracellular dd-carboxypeptidase-transpeptidase of Streptomyces R61, Biochem. J. 135:463.PubMedGoogle Scholar
  59. Galdiero, F., Tufano, M. A., and Cerciello, T. 1970, Isoelectric focusing of Clostridium botulinum Type A toxin, Arch. Mikrobiol. 74:101.PubMedCrossRefGoogle Scholar
  60. Gill, D. M., and Dinius, L. L. 1971, Observations on the structure of diphtheria toxin, J. Biol. Chem. 246:1485.PubMedGoogle Scholar
  61. Glick, J. M., Kerr, S. J., Gold, A. M., and Shemin, D. 1972, Multiple forms of colicin E3 from Escherichia coli CA-38 (col E3, col 1), Biochemistry 11:1183.PubMedCrossRefGoogle Scholar
  62. Goebel, W., and Helinski, D. R. 1971, Endonuclease I of Proteus mirabilis. I. Purification and demonstration of limited endonuclease activity, J. Biol. Chem. 246:3851.PubMedGoogle Scholar
  63. Goode, R. L., and Baldwin, J. N. 1974, Comparison of purified alpha toxins from various strains of Staphylococcus aureus, Appl. Microbiol. 28:86.PubMedGoogle Scholar
  64. Granato, P. A., and Jackson, R. W. 1971, Purification and characterization of the L component of Streptococcus zymogenes lysin, J. Bacteriol. 108:804.PubMedGoogle Scholar
  65. Haglund, H., 1971, in “Methods of Biochemical Analysis” (D. Glick, ed.), Vol. 19, pp. 1–104, Wiley (Interscience) New York.CrossRefGoogle Scholar
  66. Hassing, G. S. 1971, Partial purification and some properties of a lipase from Corynebacterium acnes, Biochim. Biophys. Acta 242:381.PubMedGoogle Scholar
  67. Hauschild, A. H. W. A., and Hilsheimer, R., 1971 Purification and characterization of the enterotoxin of Clostridium perfringens Type A, Canad. J. Microbiol. 17:1425.CrossRefGoogle Scholar
  68. Hehre, E. J., and Suzuki, H. 1966, New reactions of dextransucrase: α-d-glucosyl transfers to and from the anomeric sites of lactulose and fructose, Arch. Biochem. Biophys. 113:675.PubMedCrossRefGoogle Scholar
  69. Hetland, O., Bryn, K., and Stormer, F. C. 1971, Diacetyl (acetoin) reductase from Aerobacter aerogenes. Evidence for multiple forms of the enzyme, Eur. J. Biochem. 20:206.PubMedCrossRefGoogle Scholar
  70. Hiraoka, N., Fukumoto, J., and Tsuru, D. 1972, Studies on mold dextranases: III. Purification and some enzymatic properties of Aspergillus carneus dextranase, J. Biochem. 71:57.PubMedGoogle Scholar
  71. Howard, C. R., and Zuckerman, A. J. 1973, Electrofocusing of hepatitis B antigen, J. Gen. Virol. 20:253.PubMedCrossRefGoogle Scholar
  72. Humphryes, K. C. 1970, Isoelectric focusing of Trypanosoma brucei subgroup antigens in polyacrylamide gel thin layers. A method for resolving and characterising protein-carbohydrate complexes of an enzymic and immunological nature, J. Chromatogr. 49:503.PubMedCrossRefGoogle Scholar
  73. Hung, P. P., Robinson, H. L., and Robinson, W. 1971, Isolation and characterization of proteins from rous sarcoma virus, Virology 43:251.PubMedCrossRefGoogle Scholar
  74. Illingworth, J. A. 1972, Anomalous behavior of yeast isocitrate dehydrogenase during isoelectric focusing, Biochem. J. 129:1125.PubMedGoogle Scholar
  75. Jeansson, S., Vesterberg, O., and Wadström, T. 1972, Separation of Australia antigen by isoelectric focusing in acrylamide gel, Life Sci. 11:929.Google Scholar
  76. Jesaitis, M. A. 1970, The nature of colicin K from Proteus mirabilis, J. Exp. Med. 11:1016.CrossRefGoogle Scholar
  77. Johnson, H. N., and Debusk, A. G. 1970a, The β-galactosidase system of Neurospora crasa I. Purification and properties of the pH 4.2 enzyme, Arch. Biochem. Biophys. 138:408.CrossRefGoogle Scholar
  78. Johnson, H. N., and Debusk, A. G. 1970b, The l-galactosidase system of Neurospora crasa II. Extracellular nature of the pH 4.2 enzyme. Arch. Biochem. Biophys. 138:412.CrossRefGoogle Scholar
  79. Jolchine, G., and Reiss-Husson, F. 1972, Proteins of Rhodopseudomonas spheroides Y reaction center: Gel electrophoresis and electrofocusing studies, Biochem. Biophys. Res. Commun. 48:333.PubMedCrossRefGoogle Scholar
  80. Johnson, M., Pettersson, E., and Reinhammar, B. 1968, Isoelectric fractionation, analysis, and characterization of ampholytes in natural pH gradients. VII. The isoelectric spectra of fungal laccase A and B, Acta Chem. Scand. 22:2135.CrossRefGoogle Scholar
  81. Kanbayashi, Y., Hotta, M., and Koyama, J. 1972, Kinetic study on streptolysin O, J. Biochem. 71:227.PubMedGoogle Scholar
  82. Kassanis, B., and Kleczkowski, A. 1965, Mutual preparation of two viruses. Nature (Lond.) 205:310.CrossRefGoogle Scholar
  83. Kendal, A. P., Kiley, M. P., and Eckert, E. A. 1973, Isoelectric focusing studies of A2/1957 influenza neuraminidase and its subunits, Biochim. Biophys. Acta 317:28.PubMedGoogle Scholar
  84. Kersters-Hilderson, H., Loontiens, F. G., Claeyssens, M., and DeBruyne, C. K. 1969, Partial purification and properties of an induced β-d-xylosidase of Bacillus pumilus 12, Eur. J. Biochem. 7:434.PubMedCrossRefGoogle Scholar
  85. Kolb, E., and Harris, J. I. 1971, In search of lactate dehydrogenase from Bacillus stearothermophilus, Biochem. J. 124:76P.PubMedGoogle Scholar
  86. Kreger, A. S., and Bernheimer, A. W. 1971, Disruption of bacterial protoplasts and spheroplasts by staphylococcal delta hemolysin, Infect. Immun. 3:603.PubMedGoogle Scholar
  87. Kreger, A. S., Kim, K-S., Zaboretzky, F., and Bernheimer, A. W. 1971, Purification and properties of staphylococcal delta hemolysin, Infect. Immun. 3:449.PubMedGoogle Scholar
  88. Kreger, A. S., Cuppari, G., and Taranta, A. 1972, Isolation by electrofocusing of two lymphocyte mitogens produced by Staphylococcus aureus, Infect. Immun. 5:723.PubMedGoogle Scholar
  89. Lee, Y. C., and Wacek, V. 1970, Galactosidases from Aspergillus niger, Arch. Biochem. Biophys. 138:264.PubMedCrossRefGoogle Scholar
  90. Legler, G., von Radloff, M., and Kempfle, M. 1972, Composition, N-terminal amino acids, and chain length of a β-glucosidase from Aspergillus wentii, Biochim. Biophys. Acta 257:40.PubMedGoogle Scholar
  91. Lesk, E. M., and Blackburn, T. H. 1971, Purification of Bacteroides amylophilus protease, J. Bacteriol. 106:394.PubMedGoogle Scholar
  92. Lindstedt, G., Lindstedt, S., and Tout, M. 1970, γ-Butyrobetaine hydroxylase from Pseudomonas sp AK1, Biochemistry 9:4336.PubMedCrossRefGoogle Scholar
  93. Loach, P. A., Hadsell, R. M., Sekura, D. L., and Sterner, A. 1970, Quantitative dissolution of the membrane and preparation of photoreceptor subunits from Rhodospirillum rubrum, Biochemistry 9:3127.PubMedCrossRefGoogle Scholar
  94. Malthe-Sorenssen, D., and Stormer, F. C. 1970, The pH 6 acetolactate-forming enzyme from Serratia marcescens. Purification and properties, Eur. J. Biochem. 14:127.PubMedCrossRefGoogle Scholar
  95. Mann, K. G., Castellino, F. J., and Hargrave P. A. 1970, Molecular weight and subunit structure of yeast enolase. Biochemistry, 9:4002.PubMedCrossRefGoogle Scholar
  96. Margolis, J., and Kenrick, K. G. 1969, Two-dimensional resolution of plasma proteins by combination of polyacrylamide disc and gel gradient electrophoresis, Nature (Lond.) 221:1056.CrossRefGoogle Scholar
  97. Matthew, M., and Hedges, R. W. 1976, Analytical isoelectric focusing of R-factor determined 1-lactamases: Correlation with plasmid compatibility, J. Bacteriol. 125:713.PubMedGoogle Scholar
  98. McIver, J., Grady, G. F., and Kensch, G. T. 1975, Production and Characterization of exotoxin(s) of Shigella dysenteriae type 1, J. Infect. Dis. 131:559.PubMedCrossRefGoogle Scholar
  99. McNiven, A. C., Owen, P., and Arbuthnott, J. P. 1972, Multiple forms of staphylococcal alpha-toxin, J. Med. Microbiol. 5:113.PubMedCrossRefGoogle Scholar
  100. McQuade, A. B., and Crewther, W. G. 1969, Activity against a synthetic substrate by a preparation of extracellular proteinase from Serratia marcescens, Biochim. Biophys. Acta 191:762.PubMedGoogle Scholar
  101. Meachum, Z. D., Jr., Colisnn. H. J., Jr., and Braymer, H. D. 1971, Chemical and physical studies of Neurospora crasa invertase. Molecular weight, amino acid and carbohydrate composition, and quaternary structure, Biochemistry 10:326.PubMedCrossRefGoogle Scholar
  102. Melish, M. E., Glasgow, L. A., Turner, M. D., and Lillibridge, C. B. 1974, The staphylococcal epidermolytic toxin: Its isolation, characterization and site of action, in “Recent Advances in Staphylococcal Research” (W. W. Yotis, ed.), Ann. N. Y. Acad. Sci. 236:317.Google Scholar
  103. Metzger, J. F., Johnson, A. D., and Collins, W. S. 1972, Fractionation purification of Staphylococcus aureus enterotoxin B by electrofocusing, Biochim. Biophys. Acta 257:183.PubMedGoogle Scholar
  104. Meyer, T. E., Bartsch, R. G., and Kamen, M. D. 1971, Cytochrome c3. A class of electron transfer heme proteins found in both photosynthetic and sulfate-reducing bacteria, Biochim. Biophys. Acta 245:453.PubMedCrossRefGoogle Scholar
  105. Miller, R. L., Ramsey, G. A., Krenitsky, T. A., and Elion, G. B. 1972, Guanine phosphoribosyltransferase from Escherichia coli, specificity and properties, Biochemistry 11:4723.PubMedCrossRefGoogle Scholar
  106. Mishra, N. C., and Tatum, E. L 1970, Phosphoglucomutase mutants of Neurospora sitophila and their relation to morphology, Proc. Nat. Acad. Sci. 66:638.PubMedCrossRefGoogle Scholar
  107. Mitsui, K., Mitsui, N., and Hase, J. 1973, Clostridium perfringens exotoxins: II. Purification and some properties of theta toxin, Jap. J. Exp. Med. 43:377.PubMedGoogle Scholar
  108. Mo, Y., Harris, B. G., and Gracy, R. W. 1973, Triosephosphate isomerases and aldolases from light and dark-grown Euglena gracilis, Arch. Biochem. Biophys. 157:580.PubMedCrossRefGoogle Scholar
  109. Mollby, R., and Wadström, T. 1971, Separation of gamma hemolysin from Staphylococcus aureus, Smith 5 R, Infect. Immun. 3:633.PubMedGoogle Scholar
  110. Mollby, R., Wadström, T., Smyth, C. F., and Thelestam, M. 1974, The interaction of phospholipase C from Staphylococcus aureus and Clostridium perfringens with cell membranes, J. Hyg. Epidemiol. Microbiol. Immunol. 18:259.PubMedGoogle Scholar
  111. Nakayama, H., Okubo, S., and Takagi, Y. 1971, Repair of ultraviolet-damaged DNA in Micrococcus lysodeikticus. I. An endonuclase specific for ultraviolet-irradiated DNA, Biochim. Biophys. Acta 228:67.PubMedGoogle Scholar
  112. Nishimune, T., and Hayashi, R. 1973, Purification and some properties of thiamine binding protein of Escherichia coli, Biochim. Biophys. Acta 328:124.PubMedGoogle Scholar
  113. Oroszlan, S., Foreman, C., Kelloff, G., and Gilden, R. V. 1971, The group specific antigen and other structural proteins of hamster and mouse C-type viruses, Virology 43:665.PubMedCrossRefGoogle Scholar
  114. Pettersson, U., Philipson, L., and Hoglund, S. 1968, Structural proteins of adenoviruses. II. Purification and characterization of the adenovirus type 2 fiber antigen, Virology 35:204.PubMedCrossRefGoogle Scholar
  115. Radola, B. J. 1973, Isoelectric focusing in layers of granulated gels. I. Thin layer isoelectric focusing of proteins, Biochim. Biophys. Acta 295:412.PubMedGoogle Scholar
  116. Rebeyrotte, P., and Labbe, J.-P. 1970, Display of immunological characters common for fungal and bacterial proteases, Bull. Soc. Chim. Biol. 52:1525.PubMedGoogle Scholar
  117. Rebeyrotte, P., and Labbé, J. P. 1972, Extraction and purification of leucine aminopeptidase (EC in Aspergillus and Streptomyces proteases by ionic focusing. Comparison with pig kidney leucine amino peptidase C. R. Acad. Sci. Paris 274:2370.Google Scholar
  118. Reyes, F., and Byrde, R. J. W. 1973, Partial purification and properties of a α-N-acetylglucosaminidase from the fungus Schlerotinia fructigena, Biochem. J. 131:381.PubMedGoogle Scholar
  119. Rice, R. H., and Horst, J. 1972, Isoelectric focusing of viruses in polyacrylamide gels, Virology 49:602.PubMedCrossRefGoogle Scholar
  120. Righetti, P., and Drysdale, J. W. 1971, Isoelectric focusing in polyacrylamide gels, Biochem. Biophys. Acta 293:17.Google Scholar
  121. Righetti, P. G., and Drysdale, J. W. 1973, Small scale fractionation of proteins and nucleic acids by isoelectric focusing in polyacrylamide gels, Ann. N. Y. Acad. Sci. 209:163.PubMedCrossRefGoogle Scholar
  122. Righetti, P. G., and Drysdale, J. W. 1974, Isoelectric focusing in gels, J. Chromatogr. 98:271.PubMedCrossRefGoogle Scholar
  123. Riley, R. F., and Coleman, M. K. 1968, Isoelectric fractionation of proteins on a microscale in polyacrylamide and agarose matrices, J. Lab. Clin. Med. 72:714.PubMedGoogle Scholar
  124. Rittenhouse, H. G., Heptinstall, J., and McFadden, B. A. 1971, A hydrophobic protein from the cell envelope of Hydrogenomonas facilis, Biochemistry 10:4045.PubMedCrossRefGoogle Scholar
  125. Robb, F., Hutchinson, M. A., and Belser, W. L 1971, Anthranilate synthetase. Some physical and kinetic properties of the enzyme from Serratia marcescens, J. Biol. Chem. 246:6908.PubMedGoogle Scholar
  126. Robern, H., Stavric, S., and Dickie, N. 1974, Fractionation of staphylococcal enterotoxin A by isoelectric focusing, Experientia 30:1087.PubMedCrossRefGoogle Scholar
  127. Roberts, J., Holcenberg, J. S., and Dolowy, W. E. 1972, Isolation, crystallization and properties of Achromobacteraceae glutaminase-asparaginase with antitumor activity, J. Biol. Chem. 247:84.PubMedGoogle Scholar
  128. Rosen, B. P. 1973, Basic amino acid transport in Escherichia coli. II. Purification and properties of an arginine specific binding protein, J. Biol. Chem. 248:1211.PubMedGoogle Scholar
  129. Sakaguchi, G., Uemura, T., and Riemann, H. P. 1973, Simplified method for purification of Clostridium perfringens type A enterotoxin, Appl. Microbiol. 26:762.PubMedGoogle Scholar
  130. Sawada, T., and Sato, K. 1969, Studies on skin test antigen FST for immunodiagnosis of filariasis. III. Separation and characterization of FST3-1 by isoelectric focusing technique, Jap. J. Exp. Med. 39:533.PubMedGoogle Scholar
  131. Sawada, T., Sato, K., and Sato, S. 1969, Studies on the purification of skin test antigen SST for the diagnosis of Schistosomiasis japonica. I. The characterization and purification of antigen SST by zone electrophoresis and gel-filtration technique, Jap. J. Exp. Med. 39:339.PubMedGoogle Scholar
  132. Schaeg, W., Bingol, R., and Blobel, H. 1972, Purification of penicillinase (β-lactamase) and acid phosphatase from Staphylococcus aureus in one procedure, Biochim. Biophys. Acta 268:542.PubMedGoogle Scholar
  133. Schantz, E. J., Roessler, W. G., Woodburn, M. J., Lynch, J. M., Jacoby, H. M., Silverman, S. J., Gorman, J. C., and Spero, L. 1972, Purification and some chemical and physical properties of staphylococcal enterotoxin A, Biochemistry 11:360.PubMedCrossRefGoogle Scholar
  134. Scott, W. A., and Abramsky, T. 1973, Neurospora 6-phosphogluconate dehydrogenase. 1. Purification and characterization of the wild type enzyme, J. Biol. Chem. 248:3535.PubMedGoogle Scholar
  135. Seravalli, E., and Taranta, A. 1974, Lymphocyte transformation and macrophage migration inhibition by electrofocused and gel-filtered fractions of group A streptococcal filtrate, Cell. Immunol. 14:366.PubMedCrossRefGoogle Scholar
  136. Sheys, G. H., Arnold, W. J., Watson, J. A., Hayashi, J. A., and Doughty, C. C. 1971, Aldose reductase from Torula. I. Purification and properties, J. Biol. Chem. 246:3824.PubMedGoogle Scholar
  137. Six, H. R., and Harshman, S. 1973, Physical and chemical studies on staphylococcal α-toxins A and B, Biochemistry 12:2677.PubMedCrossRefGoogle Scholar
  138. Skjelkvale, R., and Duncan, C. L. 1975, Characterization of enterotoxin purified from Clostridium perfringens type C, Infect. Immun. 11:1061.PubMedGoogle Scholar
  139. Sletten, K., Dus, K., De Kierk, H., and Kamen, M. D. 1968, Cytochrome c2 of Rhodospirillum rubrum. I. Molecular properties of the protein and amino acid sequences of its peptides derived by the action of trypsin and thermolysin, J. Biol. Chem. 243:5492.PubMedGoogle Scholar
  140. Smyth, C. F. 1975, The identification and purification of multiple forms of theta hemolysin (theta toxin) of Clostridium perfringens type A, J. Gen. Microbiol. 87:219.PubMedGoogle Scholar
  141. Smyth, C. J., and Arbuthnott, J. P. 1972. Characteristics of Clostridium perfringens Type A α-toxin purified by isoelectric focusing, J. Gen. Microbiol. 71:11.Google Scholar
  142. Soderlind, O., Mollby, R., and Wadström, T. 1974, Purification and properties of a heat-labile enterotoxin from Escherlchia cola, Zbt. Bakt. Hyg., I. Abt. Orig. A. 229:190.Google Scholar
  143. Spero, L., Warren, J. R., and Metzger, J. 1974, Microheterogeneity of staphylococcal enterotoxin B, Biochim. Biophys. Acta 336:79.Google Scholar
  144. Sprössler, B., Lenssen, U., and Lingens, F. 1970, The properties of chorismate mutase from Saccharomyces cerevisiaeS 288 C, Hope-Seyler’s Z. Physiol. Chem. 351:1178.CrossRefGoogle Scholar
  145. Stahl, W. L., and O'Toole, R. D. 1972, Pneumococcal neuraminidase: Purification and properties, Biochim. Biophys. Acta 268:480.PubMedGoogle Scholar
  146. Stavric, S., Robern, H., and Dickie, N. 1975, Microheterogeneity of staphylococcal enterotoxln C2, Experientia 31:145.PubMedCrossRefGoogle Scholar
  147. Stern, M. L., Phillips, A. W., and Gottlieb, A. J. 1976, Physical properties of l-asparaginase from Serratia marcescens, J. Bacteriol. 125:719.PubMedGoogle Scholar
  148. Stone, K. P., and Cummings, D. J. 1972, Comparison of the internal proteins of the T-even bacteriophages, J. Mol. Biol. 64:651.PubMedCrossRefGoogle Scholar
  149. Suginaka, H., Blumberg, M. P., and Strominger, J. L. 1972, Multiple penicillin binding components in Bacillus subtilis, Bacillus cereus, Staphylococcus aureus, and Escherichia coli, J. Biol. Chem. 247:5279.PubMedGoogle Scholar
  150. Surguchev, A. P., Surgucheva, I. G., Ermokhina, T. M., and Zaitseva, G. N. 1970, Studies on methionyl-tRNA synthethase from Bacillus brevis by isoelectric focusing, Biochim. Biophys. Acta 224:623.PubMedGoogle Scholar
  151. Svensson, H. 1961, Isoelectric fractionation, analysis, and characterization of ampholytes in natural pH gradients. I. The differential equation of solute concentrations at the steady state and its solution for simple cases, Acta Chem. Scand. 15:325.CrossRefGoogle Scholar
  152. Svensson, H. 1962, Isoelectric fractionation, analysis and characterization of ampholytes in natural pH gradients. III. Description of apparatus for electrolysis in columns stabilized by density gradients and direct determination of isoelectric points, Arch. Biochem. Biophys. Suppl. 1:132.Google Scholar
  153. Tosa, T., Sano, R., Yamomoto, K., Nakamura, M., and Chibata, I. 1972, l-Asparaginase from Proteus vulgaris. Purification, crystallization, and enzymic properties, Biochemistry 11:217.PubMedCrossRefGoogle Scholar
  154. Tronson, D. A., Ritchie, G. A. F., and Nicholas, D. J. D. 1973, Purification of c-type cytochromes from Nitrosomonas europaea, Biochim. Biophys. Acta 310:331.PubMedGoogle Scholar
  155. Tsuru, D., Hattori, A., Tsuji, H., and Fukumoto, J. 1970, Studies on mold protease. III. Some physicochemical properties and amino acid composition of Rhizopus chinensis acid protease. J. Biochem. 67:415.PubMedGoogle Scholar
  156. Vesterberg, O. 1968, Studies on extracellular proteins from Staphylococcus aureus. III. Investigations on the heterogeneity of hyaluronate lyase using the method of isoelectric focusing, Biochim. Biophys. Acta 168:218.PubMedGoogle Scholar
  157. Vesterberg, O. 1969, Synthesis and isoelectric fractionation of carrier ampholytes, Acta Chem. Scand. 23:2653.CrossRefGoogle Scholar
  158. Vesterberg, O. 1971, in “Methods in Enzymology” (W. R. Jakoby, ed.), Vol. 22, pp. 389–412, Academic Press, New York.Google Scholar
  159. Vesterberg, O. 1973, Physicochemical properties of the carrier ampholytes, and some biochemical applications, Ann. N. Y. Acad. Sci. 209:23.PubMedCrossRefGoogle Scholar
  160. Vesterberg, O., Wadström, T., Vesterberg, K., Svensson, H., and Malmgren, B. 1967, Studies on extracellular proteins of Staphylococcus aureus. I. Separation and characterization of enzymes and toxins by isoelectric focusing, Biochem. Biophys. Acta 133:435.PubMedGoogle Scholar
  161. Vesterberg, K., and Vesterberg, O. 1972, Studies on staphylokinase, J. Med. Microbiol. 5:441.PubMedCrossRefGoogle Scholar
  162. Voellmy, R., and Leisinger, T. 1975, Dual role for N-acetylomithine-5-aminotransferase from Pseudomonas aeruginosa in arginine biosynthesis and arginine catabolism, J. Bacteriol. 122:799.PubMedGoogle Scholar
  163. Wadström, T. 1967, Studies on extracellular proteins from Staphylococcus aureus. II. Separation of deoxyribonucleases by isoelectric focusing. Purification and properties of the enzymes, Biochim. Biophys. Acta 147:441.PubMedGoogle Scholar
  164. Wadström, T. 1968, Studies on extracellular proteins from Staphylococcus aureus. IV. Separation of α-toxin by isoelectric focusing, Biochim. Biophys. Acta 168:228.PubMedGoogle Scholar
  165. Wadström, T. 1974, Biological properties of extracellular proteins from Staphylococcus aureus, in “Recent Advances in Staphylococcal Research” (W. W. Yotis, ed.), Ann. N.Y. Acad. Sci. 236:343.Google Scholar
  166. Wadström, T., and Hisatsune, K. 1970, Bacteriolytic enzymes from Staphylococcus aureus. Purification of an endo-l-N-acetylglucosaminidase. Biochem. J. 120:725.PubMedGoogle Scholar
  167. Wadström, T., and Möllby, R. 1971, Studies on extracellular proteins from Staphylococcus aureus. VI. Production and purification of β-haemolysin in large scale, Biochim. Biophys. Acta 242:288.PubMedGoogle Scholar
  168. Wadström, T., and Vesterberg, O. 1971, Studies on endo-β-N-acetylglucosaminidase, staphylolytic peptidase, and N-acetylmuramyl-l-alanine amidase in lysostaphin and from Staphylococcus aureus. Acta Patho. Microbiol. Scand. 79:248.Google Scholar
  169. Watanabe, M., and Kato, I. 1974, Purification and some properties of Staphylococcal alpha toxin, Jap. J. Exp. Med. 44:165.PubMedGoogle Scholar
  170. Weiner, J. H., and Hepper, L. A. 1971, A binding protein for glutamine and its relation to active transport in Escherichia coli, J. Biol. Chem. 246:6933.Google Scholar
  171. Weller, D. L, and Sjogren, R. E. 1970, Chemical heterogeneity of ribosomal protein of Escherichia coli examined by electrofocusing, Biochim. Biophys. Acta 200:508.PubMedGoogle Scholar
  172. Williamson, A. R., Solamon, M. R., and Kreth, H. W. 1973, Microheterogeneity and allomorphism of proteins, Ann. N.Y. Acad. Sci. 209:210.PubMedCrossRefGoogle Scholar
  173. Wood, T. M. 1971, The cellulase of Fusarium solani, purification and specificity of the β-(1→4)-glucanase and the β-d-glucosidase components, Biochem. J. 121:353.PubMedGoogle Scholar
  174. Wood, T. M., and McCrae, S. I. 1972, The purification and properties of the C1 component of Trichoderma koningii cellulase, Biochem. J. 128:1183.PubMedGoogle Scholar
  175. Wretlind, G., Mollby, R., and Wadström, T. 1971, Separation of two hemolysins from Aeromonas hydrophila by isoelectric focusing, Infect. Immun. 4:503.PubMedGoogle Scholar
  176. Wrigley, C. W. 1968, Gel electrofocusing—A technique for analysing multiple protein samples by isoelectric focusing, Sci. Tools 15:17.Google Scholar
  177. Yamasaki, H., and Moriyama, T. 1971, Purification, general properties and two other catalytic activities of α-ketoglutarate: Glyoxylate carboligase of Mycobacterium phlei, Biochim. Biophys. Acta 242:637.PubMedGoogle Scholar
  178. Yang, K. H., and Sugiyama, H. 1975, Purification and properties of Clostridium botulinum type F toxin, Appl. Microbiol. 29:598.PubMedGoogle Scholar
  179. Yotis, W. W., and Catsimpoolas, N. 1975, Scanning isoelectric focusing and isotachophoresis of Clostridium perfringens Type A enterotoxin, J. Appl. Bacteriol. 39:147.PubMedCrossRefGoogle Scholar
  180. Yotis, W. W., and Catsimpoolas, N. 1976, Scanning isoelectric focusing of tetanus and diptheria toxoids, Absts. Ann. Meeting Amer Soc. Microbiol. p. 203.Google Scholar
  181. Yotis, W. W., Catsimpoolas, N., Bergdoll, M. S., and Schantz, E. J. 1974, Scanning density isoelectric focusing of staphylococcal enterotoxins B and C1, Infect. Immun. 9:974.PubMedGoogle Scholar
  182. Yotis, W. W., Catsimpoolas, N., and Montiel, F. 1975, Scanning isoelectric focusing of cholera enterotoxin in polyacrylamide gels, Antonie van Leeuwenhoek J. Microbiol. Serol. 41:69.CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1977

Authors and Affiliations

  • William W. Yotis
    • 1
  1. 1.Department of Microbiology, Stritch School of MedicineLoyola University of ChicagoMaywood

Personalised recommendations