RRC ID |
39816
|
著者 |
Matsushima-Hibiya Y, Watanabe M, Kono T, Kanazawa T, Koyama K, Sugimura T, Wakabayashi K.
|
タイトル |
Purification and cloning of pierisin-2, an apoptosis-inducing protein from the cabbage butterfly, Pieris brassicae.
|
ジャーナル |
Eur J Biochem
|
Abstract |
The cabbage butterfly Pieris rapae contains a strong apoptosis-inducing substance, pierisin, against human cancer cell lines, which is thought to act via ADP-ribosylation. Here we report the purification and cloning of an apoptosis-inducing substance, designated as pierisin-2, from another cabbage butterfly, Pieris brassicae. Pierisin-2 was purified from pupae by sequential chromatography and its cytotoxic and apoptosis-inducing activities to various cancer cells were similar to those of pierisin, designated as pierisin-1, from P. rapae. cDNA cloning of pierisin-2 was performed on the basis of the partial amino-acid sequence. The nucleotide sequence indicated that the cDNA encodes an 850-amino-acid protein with a calculated molecular mass of 97 986. The deduced amino-acid sequence of pierisin-2 was 91% identical with that of pierisin-1. In vitro expressed protein in the reticulocyte lysate exhibited apoptosis-inducing activities against human gastric carcinoma TMK-1 and cervical carcinoma HeLa cells, similar to the purified native pierisin-2 from the pupae. Pierisin-2 shows regional sequence similarities with certain ADP-ribosylating toxins such as the A-subunit of cholera toxin. The results from site-directed mutagenesis at Glu165, a conserved residue among ADP-ribosylating enzymes necessary for NAD binding, and from experiments with ADP-ribosylating enzyme inhibitors suggested that pierisin-2 could be considered as an ADP-ribosylating toxin like pierisin-1.
|
巻・号 |
267(18)
|
ページ |
5742-50
|
公開日 |
2000-9-1
|
DOI |
10.1046/j.1432-1327.2000.01640.x
|
PII |
ejb1640
|
PMID |
10971585
|
MeSH |
ADP Ribose Transferases
Amino Acid Sequence
Animals
Apoptosis* / drug effects
Base Sequence
Benzamides / pharmacology
Butterflies / chemistry
Cloning, Molecular
DNA, Complementary / metabolism
Electrophoresis, Polyacrylamide Gel
HeLa Cells
Humans
Inhibitory Concentration 50
Insect Proteins / chemistry
Insect Proteins / genetics
Insect Proteins / isolation & purification*
Insect Proteins / metabolism
Insect Proteins / pharmacology
Mice
Microscopy, Fluorescence
Molecular Sequence Data
Mutagenesis, Site-Directed
Poly(ADP-ribose) Polymerase Inhibitors
Sequence Homology, Amino Acid
Species Specificity
Tumor Cells, Cultured
U937 Cells
|
引用数 |
30
|
WOS 分野
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
リソース情報 |
ヒト・動物細胞 |
HeLa(RCB0007) |