In:
Proceedings of the National Academy of Sciences, Proceedings of the National Academy of Sciences, Vol. 98, No. 5 ( 2001-02-27), p. 2226-2231
Abstract:
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.
Type of Medium:
Online Resource
ISSN:
0027-8424
,
1091-6490
DOI:
10.1073/pnas.051628898
Language:
English
Publisher:
Proceedings of the National Academy of Sciences
Publication Date:
2001
detail.hit.zdb_id:
209104-5
detail.hit.zdb_id:
1461794-8
SSG:
11
SSG:
12
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