Skip to main content
SpringerLink
Log in

Isoelectric Focusing of Microbial Proteins

  • Chapter
Biological and Biomedical Applications of Isoelectric Focusing

Part of the book series: Biological Separations ((BIOSEP))

  • 53 Accesses

Abstract

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.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
eBook
USD 16.99
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 16.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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 

  • Arbuthnott, J. P., and Beeley, J. A., 1975, “Isoelectric Focusing,“ Butterworths, London.

    Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Barret, T., and Gould, H. J. 1973, Tissue and species specificity of non-histone chromatin proteins, Biochim. Biophys. Acta 294:165.

    Google Scholar 

  • Beers, W. H., and Reich, E. 1969, Isolation and characterization of Clostridium botulinum Type B toxin, J. Biol. Chem. 244:4473.

    PubMed  CAS  Google Scholar 

  • 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 

  • Birken, S., and Pisano, M. A. 1976, Purification and properties of a polyol dehydrogenase from Cephalosporium chrysogenum, J. Bacteriol. 125:225.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Boethling, R. S. 1975, Purification and properties of a serine protease from Pseudomonas maltophilia, J. Bacteriol. 121:933.

    PubMed  CAS  Google Scholar 

  • Bosman, H. B. 1973, Protein catabolism. II. Identification of neutral and acidic proteolytic enzymes in Aspergillus niger, Biochim. Biophys. Acta 293:476.

    Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • Campbell, H. A., and Mashburn, L. T. 1969, l-asparaginase EC-2 from Escherichia coli. Some substrate specificity characteristics, Biochemistry 8:3768.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    CAS  Google Scholar 

  • Carlsson, J. 1970, A levansucrase from Streptococcus mutans, Caries Res. 4:97.

    Article  PubMed  CAS  Google Scholar 

  • Carlsson, J., Newbrun, E., and Krasse, B. 1969, Purification and properties of dextransucrase from Streptococcus sanguis, Arch. Oral. Biol. 14:469.

    Article  PubMed  CAS  Google Scholar 

  • Catsimpoolas, N. 1969, Immunoelectrofocusing in agarose gels, Clin. Chim. Acta 23:237.

    Article  PubMed  CAS  Google Scholar 

  • Catsimpoolas, N. 1970, Isoelectrc focusing of proteins in gel media, Sep. Sci. 5:523.

    Article  CAS  Google Scholar 

  • Catsimpoolas, N. 1971, Analytical scanning isoelectrofocusing: Design and operation of an in situ scanning apparatus, Anal. Biochem. 44:411.

    Article  PubMed  CAS  Google Scholar 

  • Catsimpoolas, N. 1973a, Isoelectrc focusing and isotachophoresis of proteins, Sep. Sci. 8:71.

    Article  CAS  Google Scholar 

  • Catsimpoolas, N. 1973b, Isoelectric Focusing and Isotachophoresis, Ann. N. Y. Acad. Sci. 209:1.

    Google Scholar 

  • Catsimpoolas, N. 1973c, Immunoelectrofocusing, Ann. N.Y. Acad. Sci. 209:144.

    Article  CAS  Google Scholar 

  • 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 

  • Catsimpoolas, N. 1975, Isoelectrc focusing: Fundamental aspects, Sep. Sci. 10:55.

    Article  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • Chang, P. C., and Dickie, N. 1971, Fractionation of staphylococcal enterotoxin B by isoelectric focusing, Biochim. Biophys. Acta 236:367.

    PubMed  CAS  Google Scholar 

  • Chang, P. C., Yano, Y., Dighton, M., and Dickie, N. 1971, Fractionation of staphylococcal enterotoxin C2 by isoelectric focusing, Canad. J. Microbiol. 17:1367.

    Article  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  Google Scholar 

  • Clark-Walker, G. D., and Lascelles, J. 1970, Cytochrome C550 from Spirillum itersonii: Purification and some properties, Arch. Biochem. Biophys. 136:153.

    Article  PubMed  CAS  Google Scholar 

  • Colonna, W. J., Cano, F. R., and Lampen, O. J. 1975, Microheterogeneity of yeast invertase, Biochim. Biophys. Acta 236:293.

    Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Cunningham, M., and Beachey, E. H. 1975, Immunochemical properties of streptococcal M protein purified by isoelectric focusing, J. Immunol. 115:1002.

    PubMed  CAS  Google Scholar 

  • Dale, G., and Latner, A. L. 1969, Isoelectric focusing of serum proteins in acrylamide gels followed by electrophoresis, Clin. Chim. Acta 24:61.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    CAS  Google Scholar 

  • Delisle, G., and Milazzo, F. H. 1970, The isolation of arylsulphatase isoenzymes from Pseudomonas aeruginosa, Biochim. Biophys. Acta 212:505.

    PubMed  CAS  Google Scholar 

  • Dickie, N., Yano, Y., Robern, H., and Stauric, S. 1972, On the heterogeneity of staphylococcal enterotoxin C2, Canad. J. Microbiol. 18:801.

    Article  CAS  Google Scholar 

  • Domagk, G. F., Doering, M. K., and Chilla, R. 1973, Purification and properties of ribosephosphate isomerase from Candida utilis, Eur. J. Biochem. 38:259.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • Epstein, I., and Grossowicz, N. 1975, Purification and properties of glutamate dehydrogenase from a thermophilic bacillus, J. Bacteriol. 122:1257.

    PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Fawcett, J. S. 1973, Continuous-flow isoelectric focusing and isotachophoresis, Ann. N.Y. Acad. Sci. 209:112.

    Article  PubMed  CAS  Google Scholar 

  • 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 

  • Finkelstein, R. A., and LoSpalluto, J. J. 1970, Production of highly purified choleragen and choleragenoid, J. Infect. Dis. 121(Suppl.): S63.

    Article  CAS  Google Scholar 

  • Finlayson, G. R., and Chrambach, A. 1971, Isoelectric focusing in polyacrylamide gel and its preparative application, Anal. Biochem. 40:292.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • Galdiero, F., Tufano, M. A., and Cerciello, T. 1970, Isoelectric focusing of Clostridium botulinum Type A toxin, Arch. Mikrobiol. 74:101.

    Article  PubMed  CAS  Google Scholar 

  • Gill, D. M., and Dinius, L. L. 1971, Observations on the structure of diphtheria toxin, J. Biol. Chem. 246:1485.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • Goode, R. L., and Baldwin, J. N. 1974, Comparison of purified alpha toxins from various strains of Staphylococcus aureus, Appl. Microbiol. 28:86.

    PubMed  CAS  Google Scholar 

  • Granato, P. A., and Jackson, R. W. 1971, Purification and characterization of the L component of Streptococcus zymogenes lysin, J. Bacteriol. 108:804.

    PubMed  CAS  Google Scholar 

  • Haglund, H., 1971, in “Methods of Biochemical Analysis” (D. Glick, ed.), Vol. 19, pp. 1–104, Wiley (Interscience) New York.

    Chapter  Google Scholar 

  • Hassing, G. S. 1971, Partial purification and some properties of a lipase from Corynebacterium acnes, Biochim. Biophys. Acta 242:381.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • Howard, C. R., and Zuckerman, A. J. 1973, Electrofocusing of hepatitis B antigen, J. Gen. Virol. 20:253.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Hung, P. P., Robinson, H. L., and Robinson, W. 1971, Isolation and characterization of proteins from rous sarcoma virus, Virology 43:251.

    Article  PubMed  CAS  Google Scholar 

  • Illingworth, J. A. 1972, Anomalous behavior of yeast isocitrate dehydrogenase during isoelectric focusing, Biochem. J. 129:1125.

    PubMed  CAS  Google Scholar 

  • Jeansson, S., Vesterberg, O., and Wadström, T. 1972, Separation of Australia antigen by isoelectric focusing in acrylamide gel, Life Sci. 11:929.

    CAS  Google Scholar 

  • Jesaitis, M. A. 1970, The nature of colicin K from Proteus mirabilis, J. Exp. Med. 11:1016.

    Article  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  Google Scholar 

  • Kanbayashi, Y., Hotta, M., and Koyama, J. 1972, Kinetic study on streptolysin O, J. Biochem. 71:227.

    PubMed  CAS  Google Scholar 

  • Kassanis, B., and Kleczkowski, A. 1965, Mutual preparation of two viruses. Nature (Lond.) 205:310.

    Article  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Kolb, E., and Harris, J. I. 1971, In search of lactate dehydrogenase from Bacillus stearothermophilus, Biochem. J. 124:76P.

    PubMed  CAS  Google Scholar 

  • Kreger, A. S., and Bernheimer, A. W. 1971, Disruption of bacterial protoplasts and spheroplasts by staphylococcal delta hemolysin, Infect. Immun. 3:603.

    PubMed  CAS  Google Scholar 

  • Kreger, A. S., Kim, K-S., Zaboretzky, F., and Bernheimer, A. W. 1971, Purification and properties of staphylococcal delta hemolysin, Infect. Immun. 3:449.

    PubMed  CAS  Google Scholar 

  • Kreger, A. S., Cuppari, G., and Taranta, A. 1972, Isolation by electrofocusing of two lymphocyte mitogens produced by Staphylococcus aureus, Infect. Immun. 5:723.

    PubMed  CAS  Google Scholar 

  • Lee, Y. C., and Wacek, V. 1970, Galactosidases from Aspergillus niger, Arch. Biochem. Biophys. 138:264.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • Lesk, E. M., and Blackburn, T. H. 1971, Purification of Bacteroides amylophilus protease, J. Bacteriol. 106:394.

    PubMed  CAS  Google Scholar 

  • Lindstedt, G., Lindstedt, S., and Tout, M. 1970, γ-Butyrobetaine hydroxylase from Pseudomonas sp AK1, Biochemistry 9:4336.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Mann, K. G., Castellino, F. J., and Hargrave P. A. 1970, Molecular weight and subunit structure of yeast enolase. Biochemistry, 9:4002.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • Matthew, M., and Hedges, R. W. 1976, Analytical isoelectric focusing of R-factor determined 1-lactamases: Correlation with plasmid compatibility, J. Bacteriol. 125:713.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • McNiven, A. C., Owen, P., and Arbuthnott, J. P. 1972, Multiple forms of staphylococcal alpha-toxin, J. Med. Microbiol. 5:113.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  Google Scholar 

  • 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 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Mishra, N. C., and Tatum, E. L 1970, Phosphoglucomutase mutants of Neurospora sitophila and their relation to morphology, Proc. Nat. Acad. Sci. 66:638.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Mollby, R., and Wadström, T. 1971, Separation of gamma hemolysin from Staphylococcus aureus, Smith 5 R, Infect. Immun. 3:633.

    PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • Nishimune, T., and Hayashi, R. 1973, Purification and some properties of thiamine binding protein of Escherichia coli, Biochim. Biophys. Acta 328:124.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Radola, B. J. 1973, Isoelectric focusing in layers of granulated gels. I. Thin layer isoelectric focusing of proteins, Biochim. Biophys. Acta 295:412.

    PubMed  CAS  Google Scholar 

  • Rebeyrotte, P., and Labbe, J.-P. 1970, Display of immunological characters common for fungal and bacterial proteases, Bull. Soc. Chim. Biol. 52:1525.

    PubMed  CAS  Google Scholar 

  • Rebeyrotte, P., and Labbé, J. P. 1972, Extraction and purification of leucine aminopeptidase (EC 3.4.1.1) in Aspergillus and Streptomyces proteases by ionic focusing. Comparison with pig kidney leucine amino peptidase C. R. Acad. Sci. Paris 274:2370.

    CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • Rice, R. H., and Horst, J. 1972, Isoelectric focusing of viruses in polyacrylamide gels, Virology 49:602.

    Article  PubMed  CAS  Google Scholar 

  • Righetti, P., and Drysdale, J. W. 1971, Isoelectric focusing in polyacrylamide gels, Biochem. Biophys. Acta 293:17.

    Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Righetti, P. G., and Drysdale, J. W. 1974, Isoelectric focusing in gels, J. Chromatogr. 98:271.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • Rittenhouse, H. G., Heptinstall, J., and McFadden, B. A. 1971, A hydrophobic protein from the cell envelope of Hydrogenomonas facilis, Biochemistry 10:4045.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • Robern, H., Stavric, S., and Dickie, N. 1974, Fractionation of staphylococcal enterotoxin A by isoelectric focusing, Experientia 30:1087.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • Sakaguchi, G., Uemura, T., and Riemann, H. P. 1973, Simplified method for purification of Clostridium perfringens type A enterotoxin, Appl. Microbiol. 26:762.

    PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • Six, H. R., and Harshman, S. 1973, Physical and chemical studies on staphylococcal α-toxins A and B, Biochemistry 12:2677.

    Article  PubMed  CAS  Google Scholar 

  • Skjelkvale, R., and Duncan, C. L. 1975, Characterization of enterotoxin purified from Clostridium perfringens type C, Infect. Immun. 11:1061.

    PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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 

  • 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.

    CAS  Google Scholar 

  • Spero, L., Warren, J. R., and Metzger, J. 1974, Microheterogeneity of staphylococcal enterotoxin B, Biochim. Biophys. Acta 336:79.

    CAS  Google Scholar 

  • 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.

    Article  Google Scholar 

  • Stahl, W. L., and O'Toole, R. D. 1972, Pneumococcal neuraminidase: Purification and properties, Biochim. Biophys. Acta 268:480.

    PubMed  CAS  Google Scholar 

  • Stavric, S., Robern, H., and Dickie, N. 1975, Microheterogeneity of staphylococcal enterotoxln C2, Experientia 31:145.

    Article  PubMed  CAS  Google Scholar 

  • Stern, M. L., Phillips, A. W., and Gottlieb, A. J. 1976, Physical properties of l-asparaginase from Serratia marcescens, J. Bacteriol. 125:719.

    PubMed  CAS  Google Scholar 

  • Stone, K. P., and Cummings, D. J. 1972, Comparison of the internal proteins of the T-even bacteriophages, J. Mol. Biol. 64:651.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • Vesterberg, O. 1969, Synthesis and isoelectric fractionation of carrier ampholytes, Acta Chem. Scand. 23:2653.

    Article  CAS  Google Scholar 

  • Vesterberg, O. 1971, in “Methods in Enzymology” (W. R. Jakoby, ed.), Vol. 22, pp. 389–412, Academic Press, New York.

    Google Scholar 

  • Vesterberg, O. 1973, Physicochemical properties of the carrier ampholytes, and some biochemical applications, Ann. N. Y. Acad. Sci. 209:23.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • Vesterberg, K., and Vesterberg, O. 1972, Studies on staphylokinase, J. Med. Microbiol. 5:441.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    PubMed  Google Scholar 

  • Wadström, T. 1968, Studies on extracellular proteins from Staphylococcus aureus. IV. Separation of α-toxin by isoelectric focusing, Biochim. Biophys. Acta 168:228.

    PubMed  Google Scholar 

  • 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 

  • Wadström, T., and Hisatsune, K. 1970, Bacteriolytic enzymes from Staphylococcus aureus. Purification of an endo-l-N-acetylglucosaminidase. Biochem. J. 120:725.

    PubMed  Google Scholar 

  • 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.

    PubMed  Google Scholar 

  • 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 

  • Watanabe, M., and Kato, I. 1974, Purification and some properties of Staphylococcal alpha toxin, Jap. J. Exp. Med. 44:165.

    PubMed  CAS  Google Scholar 

  • 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 

  • Weller, D. L, and Sjogren, R. E. 1970, Chemical heterogeneity of ribosomal protein of Escherichia coli examined by electrofocusing, Biochim. Biophys. Acta 200:508.

    PubMed  CAS  Google Scholar 

  • Williamson, A. R., Solamon, M. R., and Kreth, H. W. 1973, Microheterogeneity and allomorphism of proteins, Ann. N.Y. Acad. Sci. 209:210.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • Wood, T. M., and McCrae, S. I. 1972, The purification and properties of the C1 component of Trichoderma koningii cellulase, Biochem. J. 128:1183.

    PubMed  CAS  Google Scholar 

  • Wretlind, G., Mollby, R., and Wadström, T. 1971, Separation of two hemolysins from Aeromonas hydrophila by isoelectric focusing, Infect. Immun. 4:503.

    PubMed  CAS  Google Scholar 

  • Wrigley, C. W. 1968, Gel electrofocusing—A technique for analysing multiple protein samples by isoelectric focusing, Sci. Tools 15:17.

    CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • Yang, K. H., and Sugiyama, H. 1975, Purification and properties of Clostridium botulinum type F toxin, Appl. Microbiol. 29:598.

    PubMed  CAS  Google Scholar 

  • Yotis, W. W., and Catsimpoolas, N. 1975, Scanning isoelectric focusing and isotachophoresis of Clostridium perfringens Type A enterotoxin, J. Appl. Bacteriol. 39:147.

    Article  PubMed  CAS  Google Scholar 

  • 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 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Microbiology, Stritch School of Medicine, Loyola University of Chicago, Maywood, Illinois, 60153

    William W. Yotis

Authors
  1. William W. Yotis
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Massachusetts Institute of Technology, Cambridge, Massachusetts

    Nicholas Catsimpoolas

  2. Tufts University School of Medicine, Boston, Massachusetts

    James Drysdale

Rights and permissions

Reprints and permissions

Copyright information

© 1977 Plenum Press, New York

About this chapter

Cite this chapter

Yotis, W.W. (1977). Isoelectric Focusing of Microbial Proteins. In: Catsimpoolas, N., Drysdale, J. (eds) Biological and Biomedical Applications of Isoelectric Focusing. Biological Separations. Springer, Boston, MA. https://doi.org/10.1007/978-1-4613-4181-9_9

Download citation

  • DOI: https://doi.org/10.1007/978-1-4613-4181-9_9

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4613-4183-3

  • Online ISBN: 978-1-4613-4181-9

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics

Search

Navigation