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. 1967 Oct 31;126(5):923–939. doi: 10.1084/jem.126.5.923

STUDIES ON THE MODE OF ACTION OF DIPHTHERIA TOXIN

V. INHIBITION OF PEPTIDE BOND FORMATION BY TOXIN AND NAD IN CELL-FREE SYSTEMS AND ITS REVERSAL BY NICOTINAMIDE

Ronald S Goor 1, A M Pappenheimer Jr 1, Elizabeth Ames 1
PMCID: PMC2138399  PMID: 4294109

Abstract

Inhibition of soluble transferase II activity in cell-free systems by diphtheria toxin and NAD can be prevented or reversed in the presence of a sufficient concentration of nicotinamide. Quantitative studies on inhibition of peptide bond formation in cell-free extracts by toxin and NAD have indicated that two successive reversible reactions are involved. First, toxin and NAD interact mole for mole to form a relatively dissociable complex. This toxin-NAD complex then reacts with transferase II to form an enzymatically inactive product that is but slightly dissociated. In the presence of sufficient nicotinamide, however, the latter complex can be broken down to yield active transferase II once more. Based on the above model, an equation has been derived that accurately predicts the per cent inhibition of amino acid incorporation in cell-free systems at any given toxin and NAD level. The observed inhibition appears to be independent of the sensitivity to toxin of the cell species from which the extracts were derived, and depends only on the toxin and NAD concentrations. Although the model satisfactorily explains inhibition of peptide bond formation by toxin in cell-free systems, further assumptions are needed to explain how still lower concentrations of toxin are able to arrest protein synthesis completely in the living cell.

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Selected References

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  1. Bonventre P. F., Imhoff J. G. Studies on the mode of action of diphtheria toxin. I. Protein synthesis in guinea pig tissues. J Exp Med. 1966 Dec 1;124(6):1107–1122. doi: 10.1084/jem.124.6.1107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. COLLIER R. J., PAPPENHEIMER A. M., Jr STUDIES ON THE MODE OF ACTION OF DIPHTHERIA TOXIN. II. EFFECT OF TOXIN ON AMINO ACID INCORPORATION IN CELL-FREE SYSTEMS. J Exp Med. 1964 Dec 1;120:1019–1039. doi: 10.1084/jem.120.6.1019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Collier R. J. Effect of diphtheria toxin on protein synthesis: inactivation of one of the transfer factors. J Mol Biol. 1967 Apr 14;25(1):83–98. doi: 10.1016/0022-2836(67)90280-x. [DOI] [PubMed] [Google Scholar]
  4. Colombo B., Felicetti L., Baglioni C. Inhibition of protein synthesis in reticulocytes by antibiotics. I. Effects on polysomes. Biochim Biophys Acta. 1966 Apr 18;119(1):109–119. doi: 10.1016/0005-2787(66)90043-8. [DOI] [PubMed] [Google Scholar]
  5. ENNIS H. L., LUBIN M. CYCLOHEXIMIDE: ASPECTS OF INHIBITION OF PROTEIN SYNTHESIS IN MAMMALIAN CELLS. Science. 1964 Dec 11;146(3650):1474–1476. doi: 10.1126/science.146.3650.1474. [DOI] [PubMed] [Google Scholar]
  6. GABLIKS J., SOLOTOROVSKY M. Cell culture reactivity to diphtheria, Staphylococcus, tetanus and Escherichia coli toxins. J Immunol. 1962 Apr;88:505–512. [PubMed] [Google Scholar]
  7. GASIOR E., MOLDAVE K. RESOLUTION OF AMINOACYL-TRANSFERRING ENZYMES FROM RAT LIVER BY MOLECULAR SIEVE CHROMATOGRAPHY. J Biol Chem. 1965 Aug;240:3346–3352. [PubMed] [Google Scholar]
  8. GLOCK G. E., MCLEAN P. The determination of oxidized and reduced diphosphopyridine nucleotide and triphosphopyridine nucleotide in animal tissues. Biochem J. 1955 Nov;61(3):381–388. doi: 10.1042/bj0610381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. GROSSI L. G., MOLDAVE K. The effect of soluble factors on the transfer of amino acids from soluble ribonucleic acid to microsomal proteins. J Biol Chem. 1960 Aug;235:2370–2374. [PubMed] [Google Scholar]
  10. Goor R. S., Pappenheimer A. M., Jr Studies on the mode of action of diphtheria toxin. 3. Site of toxin action in cell-free extracts. J Exp Med. 1967 Nov 1;126(5):899–912. doi: 10.1084/jem.126.5.899. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Goor R. S., Pappenheimer A. M., Jr Studies on the mode of action of diphtheria toxin. IV. Specificity of the cofactor (NAD) requirement for toxin action in cell-free systems. J Exp Med. 1967 Nov 1;126(5):913–921. doi: 10.1084/jem.126.5.913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. HIEROWSKI M. INHIBITION OF PROTEIN SYNTHESIS BY CHLORTETRACYCLINE IN THE E. COLI IN VITRO SYSTEM. Proc Natl Acad Sci U S A. 1965 Mar;53:594–599. doi: 10.1073/pnas.53.3.594. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. KATO I., PAPPENHEIMER A. M., Jr An early effect of diphtheria toxin on the metabolism of mammalian cells growing in culture. J Exp Med. 1960 Aug 1;112:329–349. doi: 10.1084/jem.112.2.329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. LENNOX E. S., KAPLAN A. S. Action of diphtheria toxin on cells cultivated in vitro. Proc Soc Exp Biol Med. 1957 Aug-Sep;95(4):700–702. doi: 10.3181/00379727-95-23335. [DOI] [PubMed] [Google Scholar]
  15. NATHANS D. INHIBITION OF PROTEIN SYNTHESIS BY PUROMYCIN. Fed Proc. 1964 Sep-Oct;23:984–989. [PubMed] [Google Scholar]
  16. PAPPENHEIMER A. M., Jr, WILLIAMS C. M. The effects of diphtheria toxin on the Cecropia silkworm. J Gen Physiol. 1952 May;35(5):727–740. doi: 10.1085/jgp.35.5.727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. RAYNAUD M., BIZZINI B., RELYVELD E. H. COMPOSITION EN AMINO-ACIDES DE LA TOXINE DIPHT'ERIQUE PURIFI'EE. Bull Soc Chim Biol (Paris) 1965;47:261–266. [PubMed] [Google Scholar]
  18. Rotman B., Papermaster B. W. Membrane properties of living mammalian cells as studied by enzymatic hydrolysis of fluorogenic esters. Proc Natl Acad Sci U S A. 1966 Jan;55(1):134–141. doi: 10.1073/pnas.55.1.134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Schweet R., Heintz R. Protein synthesis. Annu Rev Biochem. 1966;35:723–758. doi: 10.1146/annurev.bi.35.070166.003451. [DOI] [PubMed] [Google Scholar]
  20. Sutter R. P., Moldave K. The interaction of aminoacyl transferase II and ribosomes. J Biol Chem. 1966 Apr 25;241(8):1698–1704. [PubMed] [Google Scholar]

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