Reference - Detail
| RRC ID | 1282 |
|---|---|
| Author | Matsushima-Hibiya Y, Watanabe M, Hidari KI, Miyamoto D, Suzuki Y, Kasama T, Kasama T, Koyama K, Sugimura T, Wakabayashi K. |
| Title | Identification of glycosphingolipid receptors for pierisin-1, a guanine-specific ADP-ribosylating toxin from the cabbage butterfly. |
| Journal | J Biol Chem |
| Abstract |
Pierisin-1, a cytotoxic protein found naturally in the cabbage butterfly, induces apoptosis of mammalian cells. Our recent studies suggest that pierisin-1 consists of an N-terminal ADP-ribosyltransferase domain, and a C-terminal region that binds to receptors on the surfaces of target cells and incorporates the protein into cells. The present study was undertaken to identify receptors for pierisin-1. The cross-linking and cloning experiments suggested that the proteins on cell membrane had no binding ability to pierisin-1. Inhibitory assays of fractionated lipids from human cervical carcinoma HeLa cells, which are highly sensitive to pierisin-1, indicated neutral glycosphingolipids on the cell surface to show receptor activity. Inhibitory assays and TLC immunostaining using anti-pierisin-1 antibodies demonstrated two neutral glycosphingolipids as active components. Analysis of their structures with glycosphingolipid-specific antibodies and negative secondary ion mass spectrometry identified them as globotriaosylceramide (Gb3) and globotetraosylceramide (Gb4). The receptor activities of Gb3 and Gb4 for pierisin-1 were also confirmed with these authentic compounds. Pierisin-1-insensitive mouse melanoma MEB4 cells were found to lack pierisin-1 receptors, including Gb3 and Gb4, but pretreatment of the cells with glycosphingolipid Gb3 or Gb4 enhanced their sensitivity to pierisin-1. Thus, Gb3 and Gb4 were proven to serve as pierisin-1 receptors. The C-terminal region of pierisin-1 consists of possible lectin domains of a ricin B-chain, containing QXW sequences, which are essential for its structural organization. Alteration of QXW by site-directed mutagenesis caused marked reduction of pierisin-1 cytotoxicity. Thus, our results suggest that pierisin-1 binds to Gb3 and Gb4 receptors at the C-terminal region, in a manner similar to ricin, and then exhibits cytotoxicity after incorporation into the cell. |
| Volume | 278(11) |
| Pages | 9972-8 |
| Published | 2003-3-14 |
| DOI | 10.1074/jbc.m212114200 |
| PII | S0021-9258(19)71393-3 |
| PMID | 12645583 |
| MeSH | ADP Ribose Transferases Amino Acid Sequence Animals Base Sequence Butterflies Chromatography, Thin Layer Cross-Linking Reagents / pharmacology Dose-Response Relationship, Drug Glycosphingolipids / metabolism* HeLa Cells Humans Inhibitory Concentration 50 Insect Proteins / chemistry* Insect Proteins / metabolism* Lectins Mass Spectrometry Mice Models, Molecular Molecular Sequence Data Mutagenesis, Site-Directed Phospholipids / metabolism Protein Binding Protein Structure, Tertiary Receptors, Cell Surface / metabolism* Sequence Homology, Amino Acid |
| IF | 4.238 |
| Times Cited | 35 |
| WOS Category | BIOCHEMISTRY & MOLECULAR BIOLOGY |
| Altmetric score |
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| The most frequently cited source | Patent(IFI CLAIMS) |
| Total number of mentions | 1 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Human and Animal Cells | HeLa(RCB0007) MEB4(RCB1027) |