Document Detail


Genetic construction and properties of a diphtheria toxin-related substance P fusion protein: in vitro destruction of cells bearing substance P receptors.
MedLine Citation:
PMID:  8692995     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We have genetically replaced the native receptor binding domain of diphtheria toxin with an extended form of substance P (SP): SP-glycine (SP-Gly). The resulting fusion protein, DAB389SP-Gly, is composed of the catalytic and transmembrane domains of diphtheria toxin genetically coupled to SP-Gly. Because native SP requires a C-terminal amide moiety to bind with high affinity to the SP receptor, the precursor form of the fusion toxin, DAB389SP-Gly, was converted to DAB389SP by treatment with peptidylglycine-alpha-amidating monooxygenase. We demonstrate that following conversion, DAB389SP is selectively cytotoxic for cell lines that express either the rat or the human SP receptor. We also demonstrate that the cytotoxic action of DAB389SP is mediated via the SP receptor and dependent upon passage through an acidic compartment. To our knowledge, this is the first reported use of a neuropeptide as the targeting ligand for a fusion toxin; and the first instance in which an inactive precursor form of a fusion toxin is converted to the active form by a posttranslational modification.
Authors:
C E Fisher; J A Sutherland; J E Krause; J R Murphy; S E Leeman; J C vanderSpek
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  93     ISSN:  0027-8424     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  1996 Jul 
Date Detail:
Created Date:  1996-08-29     Completed Date:  1996-08-29     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  7341-5     Citation Subset:  IM    
Affiliation:
Evans Department of Clinical Research, Boston University Medical Center Hospital, Boston, MA 02118, USA.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
Base Sequence
Binding, Competitive
CHO Cells
Cell Line
Cell Survival / drug effects*
Cloning, Molecular
Cricetinae
Diphtheria Toxin / biosynthesis*,  toxicity*
Escherichia coli
Humans
Kinetics
Molecular Sequence Data
Rats
Receptors, Neurokinin-1 / biosynthesis,  physiology*
Recombinant Fusion Proteins / biosynthesis,  toxicity
Substance P / analogs & derivatives*,  biosynthesis,  metabolism,  toxicity
Transfection
Tumor Cells, Cultured
Grant Support
ID/Acronym/Agency:
CA 60934/CA/NCI NIH HHS
Chemical
Reg. No./Substance:
0/Diphtheria Toxin; 0/Receptors, Neurokinin-1; 0/Recombinant Fusion Proteins; 33507-63-0/Substance P; 98093-85-7/substance P, Gly(12)-
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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