Document Detail


Distinct roles for the N- and C-terminal regions in the cytotoxicity of pierisin-1, a putative ADP-ribosylating toxin from cabbage butterfly, against mammalian cells.
MedLine Citation:
PMID:  11226221     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Pierisin-1 is an 850-aa cytotoxic protein found in the cabbage butterfly, Pieris rapae, and has been suggested to consist of an N-terminal region with ADP-ribosyltransferase domain and of a C-terminal region that might have a receptor-binding domain. To elucidate the role of each region, we investigated the functions of various fragments of pierisin-1. In vitro expressed polypeptide consisting of amino acid residues 1-233 or 234-850 of pierisin-1 alone did not show cytotoxicity against human cervical carcinoma HeLa cells. However, the presence of both polypeptides in the culture medium showed some of the original cytotoxic activity. Introduction of the N-terminal polypeptide alone by electroporation also induced cell death in HeLa cells, and even in the mouse melanoma MEB4 cells insensitive to pierisin-1. Thus, the N-terminal region has a principal role in the cytotoxicity of pierisin-1 inside mammalian cells. Analyses of incorporated pierisin-1 indicated that the entire protein, regardless of whether it consisted of a single polypeptide or two separate N- and C-terminal polypeptides, was incorporated into HeLa cells. However, neither of the terminal polypeptides was incorporated when each polypeptide was present separately. These findings indicate that the C-terminal region is important for the incorporation of pierisin-1. Moreover, presence of receptor for pierisin-1 in the lipid fraction of cell membrane was suggested. The cytotoxic effects of pierisin-1 were enhanced by previous treatment with trypsin, producing "nicked" pierisin-1. Generation of the N-terminal fragment in HeLa cells was detected after application of intact entire molecule of pierisin-1. From the above observations, it is suggested that after incorporation of pierisin-1 into the cell by interaction of its C-terminal region with the receptor in the cell membrane, the entire protein is cleaved into the N- and C-terminal fragments with intracellular protease, and the N-terminal fragment then exhibits cytotoxicity.
Authors:
T Kanazawa; M Watanabe; Y Matsushima-Hibiya; T Kono; N Tanaka; K Koyama; T Sugimura; K Wakabayashi
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  98     ISSN:  0027-8424     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  2001 Feb 
Date Detail:
Created Date:  2001-03-06     Completed Date:  2001-09-13     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:  2226-31     Citation Subset:  IM    
Affiliation:
Cancer Prevention Division, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, Japan. tkanazaw@gan2.ncc.go.jp
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
ADP Ribose Transferases
ADP-Ribosylation Factors / chemistry,  pharmacology*
Amino Acid Sequence
Animals
Apoptosis / drug effects
Base Sequence
Butterflies
Cell Survival / drug effects*
Cytotoxins / chemistry,  pharmacology*
DNA Primers
Fluorescent Dyes
Humans
Insect Proteins / chemistry,  pharmacology*
Tumor Cells, Cultured
Chemical
Reg. No./Substance:
0/Cytotoxins; 0/DNA Primers; 0/Fluorescent Dyes; 0/Insect Proteins; 0/pierisin protein, insect; EC 2.4.2.-/ADP Ribose Transferases; EC 3.6.5.2/ADP-Ribosylation Factors
Comments/Corrections

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


Previous Document:  Mutated plant lectin library useful to identify different cells.
Next Document:  The nitrite reductase from Pseudomonas aeruginosa: essential role of two active-site histidines in t...