Home LiteratureArticle Details
PMID: 3095831 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Genetic construction, expression, and melanoma-selective cytotoxicity of a diphtheria toxin-related alpha-melanocyte-stimulating hormone fusion protein.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 83 ·No. 21 ·1986-11-00 ·Pages 8258-62

Murphy JR, Bishai W, Borowski M, Miyanohara A, Boyd J, Nagle S

Abstract

The structural gene for diphtheria toxin, tox, has been modified at its Sph I site by the introduction of an oligonucleotide linker encoding a unique Pst I restriction endonuclease site and a synthetic oligonucleotide encoding alpha-melanocyte-stimulating hormone (alpha-MSH). The resulting fusion gene directs the expression of a diphtheria toxin-related alpha-MSH hybrid protein in which the diphtheria toxin receptor-binding domain has been replaced with alpha-MSH sequences. The chimeric toxin has been partially purified from periplasmic extracts of recombinant Escherichia coli K-12 and has been found to be selectively toxic for alpha-MSH receptor-positive human malignant melanoma NEL-M1 cells in vitro.

MeSH Terms
ADP Ribose Transferases Ammonium Chloride/pharmacology Cell Line Diphtheria Toxin/genetics Genes Humans Melanocyte-Stimulating Hormones/genetics Melanoma/drug therapy Molecular Weight Pentosyltransferases/analysis Recombinant Fusion Proteins/biosynthesis,genetics,pharmacology Recombinant Proteins/genetics
Chemicals
Diphtheria Toxin Recombinant Fusion Proteins Recombinant Proteins Ammonium Chloride Melanocyte-Stimulating Hormones ADP Ribose Transferases Pentosyltransferases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Murphy J R
Bishai W
Borowski M
Miyanohara A
Boyd J
Nagle S
References (40)
40 references, click to expand
  1. Hormone-receptor interactions. The message sequence of alpha-melanotropin: demonstration of two active sites.
    Clin Endocrinol (Oxf). 1976;5 Suppl:41S-48S PMID: 212233
  2. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  3. A hybrid protein containing the toxic subunit of ricin and the cell-specific subunit of human chorionic gonadotropin. I. Synthesis and characterization.
    J Biol Chem. 1979 Feb 25;254(4):1022-7 PMID: 762110
  4. Preparation of concanavalin A-ricin A-chain conjugate and its biologic activity against various cultured cells.
    J Natl Cancer Inst. 1979 Jun;62(6):1387-95 PMID: 286111
  5. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
    Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4 PMID: 388439
  6. Inhibitory effect of ammonium chloride and chloroquine on the entry of the toxic lectin modeccin into HeLa cells.
    Biochem Biophys Res Commun. 1979 Sep 27;90(2):648-55 PMID: 508328
  7. Inhibition of diphtheria toxin degradation and cytotoxic action by chloroquine.
    J Biol Chem. 1980 Mar 25;255(6):2247-50 PMID: 6766935
  8. Toxicity of diphtheria toxin-related proteins produced by suppression of nonsense mutations.
    J Biol Chem. 1980 Nov 25;255(22):10658-62 PMID: 6776122
  9. Epidermal growth factor-toxin A chain conjugates: EGF-ricin A is a potent toxin while EGF-diphtheria fragment A is nontoxic.
    Cell. 1980 Nov;22(2 Pt 2):563-70 PMID: 6256086
  10. Kinetics of adenosinediphosphoribosylation of elongation factor 2 in cells exposed to diphtheria toxin.
    Infect Immun. 1981 May;32(2):575-82 PMID: 7251138
  11. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A.
    Anal Biochem. 1981 Apr;112(2):195-203 PMID: 6266278
  12. Asialoglycoprotein receptor mediates the toxic effects of an asialofetuin-diphtheria toxin fragment A conjugate on cultured rat hepatocytes.
    Proc Natl Acad Sci U S A. 1981 Jun;78(6):3383-7 PMID: 6167984
  13. Restriction map of corynebacteriophages beta c and beta vir and physical localization of the diphtheria tox operon.
    J Bacteriol. 1981 Oct;148(1):124-30 PMID: 6270058
  14. Identification of deoxyribonucleic acid restriction fragments of beta-converting corynebacteriophages that carry the gene for diphtheria toxin.
    J Bacteriol. 1981 Oct;148(1):153-62 PMID: 6270061
  15. Entry of the toxic proteins abrin, modeccin, ricin, and diphtheria toxin into cells. II. Effect of pH, metabolic inhibitors, and ionophores and evidence for toxin penetration from endocytotic vesicles.
    J Biol Chem. 1982 Jul 10;257(13):7504-13 PMID: 7085634
  16. Isolation of hybridoma cell lines and characterization of monoclonal antibodies against cholera enterotoxin and its subunits.
    Infect Immun. 1982 Oct;38(1):267-72 PMID: 6183210
  17. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers.
    Gene. 1982 Oct;19(3):259-68 PMID: 6295879
  18. Thyrotropin-releasing hormone-diphtheria toxin-related polypeptide conjugates. Potential role of the hydrophobic domain in toxin entry.
    J Biol Chem. 1983 Feb 10;258(3):1565-70 PMID: 6296105
  19. Immunotoxins: a new approach to cancer therapy.
    Science. 1983 Feb 11;219(4585):644-50 PMID: 6218613
  20. pEMBL: a new family of single stranded plasmids.
    Nucleic Acids Res. 1983 Mar 25;11(6):1645-55 PMID: 6300771
  21. Cloned fragment A of diphtheria toxin is expressed and secreted into the periplasmic space of Escherichia coli K12.
    Science. 1983 Apr 29;220(4596):515-7 PMID: 6403984
  22. Nucleotide sequence and expression of the diphtheria tox228 gene in Escherichia coli.
    Science. 1983 Aug 26;221(4613):855-8 PMID: 6348945
  23. Molecular cloning and expression of gene fragments from corynebacteriophage beta encoding enzymatically active peptides of diphtheria toxin.
    J Bacteriol. 1983 Nov;156(2):680-5 PMID: 6313613
  24. The complete nucleotide sequence of the gene coding for diphtheria toxin in the corynephage omega (tox+) genome.
    Nucleic Acids Res. 1983 Oct 11;11(19):6589-95 PMID: 6314249
  25. Nucleotide sequence of the structural gene for diphtheria toxin carried by corynebacteriophage beta.
    Proc Natl Acad Sci U S A. 1983 Nov;80(22):6853-7 PMID: 6316330
  26. Cloned diphtheria toxin fragment A is expressed from the tox promoter and exported to the periplasm by the SecA apparatus of Escherichia coli K12.
    J Biol Chem. 1983 Dec 25;258(24):15016-20 PMID: 6317684
  27. Suppression of DNA synthesis in NEL-M1 human melanoma cells by triamcinolone acetonide.
    Cancer Res. 1985 Apr;45(4):1633-7 PMID: 3919942
  28. Production of abnormal proteins in E. coli stimulates transcription of lon and other heat shock genes.
    Cell. 1985 Jun;41(2):587-95 PMID: 3886165
  29. Synthesis and in vitro activity of a hormone-diphtheria toxin fragment A hybrid.
    Biochem Biophys Res Commun. 1985 Dec 17;133(2):430-5 PMID: 3002340
  30. INFECTION OF HUMAN AND SIMIAN TISSUE CULTURES WITH ROUS SARCOMA VIRUS.
    Proc Natl Acad Sci U S A. 1964 Jul;52:53-9 PMID: 14192657
  31. In vitro inhibition of diphtheria toxin action by ammonium salts and amines.
    J Bacteriol. 1965 Dec;90(6):1552-6 PMID: 5856535
  32. A complementation analysis of the restriction and modification of DNA in Escherichia coli.
    J Mol Biol. 1969 May 14;41(3):459-72 PMID: 4896022
  33. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  34. Genetics of somatic mammalian cells. X. Complementation analysis of glycine-requiring mutants.
    Proc Natl Acad Sci U S A. 1969 Dec;64(4):1284-91 PMID: 5271752
  35. Mutation in the structural gene for diphtheria toxin carried by temperate phage .
    Nat New Biol. 1971 Sep 1;233(35):8-11 PMID: 4999827
  36. Effect of ethylenediaminetetraacetic acid, Triton X-100, and lysozyme on the morphology and chemical composition of isolate cell walls of Escherichia coli.
    J Bacteriol. 1971 Oct;108(1):553-63 PMID: 5001205
  37. Mechanism of action of the toxic lectins abrin and ricin.
    Nature. 1974 Jun 14;249(458):627-31 PMID: 4857870
  38. Association of cell surface receptors for melanotropin with the Golgi region in mouse melanoma cells.
    Proc Natl Acad Sci U S A. 1976 Feb;73(2):559-62 PMID: 174112
  39. Artificial hybrid protein containing a toxic protein fragment and a cell membrane receptor-binding moiety in a disulfide conjugate. II. Biochemical and biologic properties of diphtheria toxin fragment A-S-S-human placental lactogen.
    J Biol Chem. 1977 Feb 25;252(4):1515-22 PMID: 190237
  40. Chimeric toxins: toxic, disulfide-linked conjugate of concanavalin A with fragment A from diphtheria toxin.
    Proc Natl Acad Sci U S A. 1978 Nov;75(11):5319-23 PMID: 281682
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.83.21.8258
Published
1986-11-00
Pages
8258-62
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC386907
Subset
IM
Grants
NIAID NIH HHS · AI-21628 · United States
NCI NIH HHS · CA-41746 · United States
External Links
PubMed source DOI: 10.1073/pnas.83.21.8258 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2