1887

Abstract

5′-nucleotidase was purified to apparent homogeneity as judged by polyacrylamide gel electrophoresis. Its molecular weight was estimated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, by gel exclusion chromatography, and by membrane filtration; values of 66000, 48500, and 15000 to 30000, respectively, were obtained. The enzyme was completely released from bacteria by osmotic shock treatment. The apparently anomalous behaviour of 5′-nucleotidase in terms of the molecular sieving hypothesis for the release of enzymes by osmotic shock proposed by Smith & Wyatt (1974) and extended by Broad & Smith (1979) is discussed.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-123-2-241
1981-04-01
2024-12-06
Loading full text...

Full text loading...

/deliver/fulltext/micro/123/2/mic-123-2-241.html?itemId=/content/journal/micro/10.1099/00221287-123-2-241&mimeType=html&fmt=ahah

References

  1. Anderson E.S., Datta N. 1965; Resistance to penicillins and its transfer in Enterobacteriaceae. Lancet 1:407–409
    [Google Scholar]
  2. Andrews P. 1964; Estimation of the molecular weights of proteins by Sephadex gel-filtration. Biochemical Journal 91:222–233
    [Google Scholar]
  3. Broad D.F., Smith J.T. 1979; Release of enzymes from bacteria. Journal of Pharmacy and Pharmacology 31:30P
    [Google Scholar]
  4. Chen P.S., Toribara T.Y., Warner H. 1956; Microdetermination of phosphorus. Analytical Chemistry 28:1757–1761
    [Google Scholar]
  5. Clowes R.C., Rowley D. 1954; Some observations on linkage effects in genetic recombination in Escherichia coli K12. Journal of General Microbiology 11:250–260
    [Google Scholar]
  6. Dale J.W. 1970 The purification and properties of penicillinases from penicillin-resistant Gramnegative bacteria, some of which harbour R-factors. Ph.D. thesis: University of London.;
    [Google Scholar]
  7. Dale J.W. 1971; Characterization of the β-lactamase specified by the resistance factor R1818 in Escherichia coli K12 and other Gram-negative bacteria. Biochemical Journal 123:501–505
    [Google Scholar]
  8. Dale J.W., Smith J.T. 1971; The purification and properties of the β-lactamase specified by the resistance factor R1818 in Escherichia coli and Proteus mirabilis . Biochemical Journal 123:493–500
    [Google Scholar]
  9. Dale J.W., Smith J.T. 1974; R.factor mediated β-lactamases that hydrolyze oxacillin: evidence for two distinct groups. Journal of Bacteriology 119:351–356
    [Google Scholar]
  10. Dale J.W., Smith J.T. 1976; The dimeric nature of an R.factor mediated β-lactamase. Biochemical and Biophysical Research Communications 68:1000–1005
    [Google Scholar]
  11. Davis B.D., Mingioli E.S. 1950; Mutants of Escherichia coli requiring methionine or vitamin B12. Journal of Bacteriology 60:17–28
    [Google Scholar]
  12. Davis B.J. 1964; Disc electrophoresis. II. Method and application to human serum proteins. Annals of the New York Academy of Sciences 121:404–427
    [Google Scholar]
  13. Dvorak H.F., Heppel L.A. 1968; Metalloenzymes released from Escherichia coli by osmotic shock. II. Evidence that 5´ nucleotidase and cyclic phosphodiesterase are zinc metalloenzymes. Journal of Biological Chemistry 243:2647–2653
    [Google Scholar]
  14. Hedges R.W., Datta N., Kontamichalou P., Smith J.T. 1974; Molecular specificities of R. factor-determined β-lactamases: correlation with plasmid compatibility. Journal of Bacteriology 117:56–62
    [Google Scholar]
  15. Heppel L.A. 1971; The concept of periplasmic enzymes. In Structure and Function of Biological Membranes pp. 223–247 Edited by Rothfield L. I. New York: Academic Press.;
    [Google Scholar]
  16. Mitchell P. 1961; Approaches to the analysis of specific membrane transport. In Biology Structure and Function, vol. 2, Proceedings of the First IUB/IUBS International Symposium pp. 581–603 Edited by Goodwin T.W., Lindberg O. London & New York: Academic Press.;
    [Google Scholar]
  17. Neu H.C. 1967a; The 5´-nucleotidase of Escherichia coli. I.Purification and properties. Journal of Biological Chemistry 242:3896–3904
    [Google Scholar]
  18. Neu H.C. 1967b; The 5´-nucleotidase of Escherichia coli. II.Surface localization and purification of 5´-nucleotidase inhibitor. Journal of Biological Chemistry 242:3905–3911
    [Google Scholar]
  19. Neu H.C., Chou J. 1967; Release of surface enzymes in Enterobacteriaceae by osmotic shock. Journal of Bacteriology 94:1934–1945
    [Google Scholar]
  20. Neu H.C., Heppel L.A. 1964; On the surface localization of enzymes in E.coli . Biochemical and Biophysical Research Communications 17:215–219
    [Google Scholar]
  21. Smith J.T., Wyatt J.M. 1974; Relation of R.factor and chromosomal β-lactamase with the periplasmic space. Journal of Bacteriology 117:931–939
    [Google Scholar]
  22. Spahr P.F., Hollingworth B.R. 1961; Purification and mechanism of action of ribonuclease from Escherichia coli ribosomes. Journal of Biological Chemistry 236:823–831
    [Google Scholar]
  23. Uyemura D., Lehman I.R. 1976; Biochemical characterization of mutant forms of DNA polymerase I from Escherichia coli. I.The polA12 mutation. Journal of Biological Chemistry 251:4078–4084
    [Google Scholar]
  24. Waddell W.J. 1956; A simple ultraviolet spectrophotometric method for the determination of protein. Journal of Laboratory and Clinical Medicine 48:311–314
    [Google Scholar]
  25. Weber K., Osborn M. 1969; The reliability of molecular weight determinations by dodecyl sulphate-polyacrylamide gel electrophoresis. Journal of Biological Chemistry 224:4406–4412
    [Google Scholar]
  26. Witchitz J.L., Chabbert Y.A. 1971; High level transferable resistance to gentamicin. Journal of Antibiotics 24:137–139
    [Google Scholar]
  27. Wyatt J.M. 1972 Episomal and chromosomal bacterial enzymes: their retention and release. Ph.D.thesis: University of London.;
    [Google Scholar]
/content/journal/micro/10.1099/00221287-123-2-241
Loading
/content/journal/micro/10.1099/00221287-123-2-241
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error