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PMID: 1978925 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Role of arginine 180 and glutamic acid 177 of ricin toxin A chain in enzymatic inactivation of ribosomes.

Molecular and cellular biology ·Vol. 10 ·No. 12 ·1990-12-00 ·Pages 6257-63

Frankel A, Welsh P, Richardson J, Robertus JD

Abstract

The gene for ricin toxin A chain was modified by site-specific mutagenesis to change arginine 180 to alanine, glutamine, methionine, lysine, or histidine. Separately, glutamic acid 177 was changed to alanine and glutamic acid 208 was changed to aspartic acid. Both the wild-type and mutant proteins were expressed in Escherichia coli and, when soluble, purified and tested quantitatively for enzyme activity. A positive charge at position 180 was found necessary for solubility of the protein and for enzyme activity. Similarly, a negative charge with a proper geometry in the vicinity of position 177 was critical for ricin toxin A chain catalysis. When glutamic acid 177 was converted to alanine, nearby glutamic acid 208 could largely substitute for it. This observation provided valuable structural information concerning the nature of second-site mutations.

MeSH Terms
Animals Arginine Escherichia coli/genetics Glutamates Glutamic Acid Kinetics Models, Molecular Mutagenesis, Site-Directed Plasmids Protein Biosynthesis/drug effects Protein Conformation Rabbits Recombinant Proteins/isolation & purification,pharmacology Reticulocytes/drug effects,metabolism Ribosomes/drug effects,metabolism Ricin/genetics,isolation & purification,pharmacology Saccharomyces cerevisiae/genetics,growth & development
Chemicals
Glutamates Recombinant Proteins Glutamic Acid Ricin Arginine
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Frankel A
Florida Hospital Cancer, Altamonte Springs.
Welsh P
Richardson J
Robertus J D
References (26)
26 references, click to expand
  1. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  2. Gene mutations in human haemoglobin: the chemical difference between normal and sickle cell haemoglobin.
    Nature. 1957 Aug 17;180(4581):326-8 PMID: 13464827
  3. Physical organic models for the mechanism of lysozyme action.
    Adv Enzymol Relat Areas Mol Biol. 1973;37:1-60 PMID: 4570067
  4. Chemical structure of a modification of the Escherichia coli ribonucleic acid polymerase alpha polypeptides induced by bacteriophage T4 infection.
    J Biol Chem. 1974 Oct 10;249(19):6181-90 PMID: 4371081
  5. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
    Anal Biochem. 1976 May 7;72:248-54 PMID: 942051
  6. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  7. Structural determinants of Ricinus communis agglutinin and toxin specificity for oligosaccharides.
    J Biol Chem. 1979 Oct 10;254(19):9795-9 PMID: 489569
  8. Entry of lethal doses of abrin, ricin and modeccin into the cytosol of HeLa cells.
    Exp Cell Res. 1980 Apr;126(2):321-6 PMID: 7363949
  9. A rapid boiling method for the preparation of bacterial plasmids.
    Anal Biochem. 1981 Jun;114(1):193-7 PMID: 6269464
  10. Transformation of intact yeast cells treated with alkali cations.
    J Bacteriol. 1983 Jan;153(1):163-8 PMID: 6336730
  11. Disuccinimidyl suberate cross-linked ricin does not inhibit cell-free protein synthesis.
    Biochem Biophys Res Commun. 1982 Nov 16;109(1):7-13 PMID: 7159430
  12. Sequences that regulate the divergent GAL1-GAL10 promoter in Saccharomyces cerevisiae.
    Mol Cell Biol. 1984 Aug;4(8):1440-8 PMID: 6092912
  13. The three-dimensional structure of ricin at 2.8 A.
    J Biol Chem. 1987 Apr 15;262(11):5398-403 PMID: 3558397
  14. Transition-state structures for N-glycoside hydrolysis of AMP by acid and by AMP nucleosidase in the presence and absence of allosteric activator.
    Biochemistry. 1987 Feb 10;26(3):921-30 PMID: 3552038
  15. The mechanism of action of ricin and related toxic lectins on eukaryotic ribosomes. The site and the characteristics of the modification in 28 S ribosomal RNA caused by the toxins.
    J Biol Chem. 1987 Apr 25;262(12):5908-12 PMID: 3571242
  16. Interaction of Cibacron blue F3GA and polynucleotides with ricin A-chain, 60 S ribosomal subunit-inactivating protein.
    Biochim Biophys Acta. 1987 Aug 5;914(2):177-84 PMID: 3607071
  17. Evidence that glutamic acid 167 is an active-site residue of Shiga-like toxin I.
    Proc Natl Acad Sci U S A. 1988 Apr;85(8):2568-72 PMID: 3357883
  18. Interaction of the bovine papillomavirus type 1 E2 transcriptional control protein with the viral enhancer: purification of the DNA-binding domain and analysis of its contact points with DNA.
    J Virol. 1988 Jun;62(6):1925-31 PMID: 2835497
  19. Ribosome-inhibiting proteins, retroviral reverse transcriptases, and RNase H share common structural elements.
    Proteins. 1988;3(1):53-9 PMID: 2453878
  20. Isolation and in vitro expression of the ricin A-chain gene: effect of deletions on biological activity.
    Nucleic Acids Res. 1989 Feb 25;17(4):1717-32 PMID: 2922291
  21. Selection and characterization of ricin toxin A-chain mutations in Saccharomyces cerevisiae.
    Mol Cell Biol. 1989 Feb;9(2):415-20 PMID: 2469000
  22. A conserved region in alpha-macroglobulins participates in binding to the mammalian alpha-macroglobulin receptor.
    Biochemistry. 1989 Feb 7;28(3):1406-12 PMID: 2469470
  23. Ribosome inactivation by ricin A chain: a sensitive method to assess the activity of wild-type and mutant polypeptides.
    EMBO J. 1989 Jan;8(1):301-8 PMID: 2714255
  24. Site-directed mutagenesis at amino terminus of recombinant ricin A chain.
    Int J Pept Protein Res. 1989 Jul;34(1):2-5 PMID: 2551831
  25. Role of glutamic acid 177 of the ricin toxin A chain in enzymatic inactivation of ribosomes.
    Mol Cell Biol. 1989 Nov;9(11):5012-21 PMID: 2689871
  26. Treatment of abrin and ricin with -mercaptoethanol opposite effects on their toxicity in mice and their ability to inhibit protein synthesis in a cell-free system.
    FEBS Lett. 1972 Nov 15;28(1):48-50 PMID: 4646874
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.10.12.6257-6263.1990
Published
1990-12-00
Pages
6257-63
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC362900
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.10.12.6257-6263.1990 PMC Full Text View journal details
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