論文 - 詳細
| RRC ID | 68079 |
|---|---|
| 著者 | Liew YJ, Soh WT, Jiemy WF, Hwang JS. |
| タイトル | Mutagenesis and functional analysis of the pore-forming toxin HALT-1 from Hydra magnipapillata. |
| ジャーナル | Toxins (Basel) |
| Abstract |
Actinoporins are small 18.5 kDa pore-forming toxins. A family of six actinoporin genes has been identified in the genome of Hydra magnipapillata, and HALT-1 (Hydra actinoporin-like toxin-1) has been shown to have haemolytic activity. In this study, we have used site-directed mutagenesis to investigate the role of amino acids in the pore-forming N-terminal region and the conserved aromatic cluster required for cell membrane binding. A total of 10 mutants of HALT-1 were constructed and tested for their haemolytic and cytolytic activity on human erythrocytes and HeLa cells, respectively. Insertion of 1-4 negatively charged residues in the N-terminal region of HALT-1 strongly reduced haemolytic and cytolytic activity, suggesting that the length or charge of the N-terminal region is critical for pore-forming activity. Moreover, substitution of amino acids in the conserved aromatic cluster reduced haemolytic and cytolytic activity by more than 80%, suggesting that these aromatic amino acids are important for attachment to the lipid membrane as shown for other actinoporins. The results suggest that HALT-1 and other actinoporins share similar mechanisms of pore formation and that it is critical for HALT-1 to maintain an amphipathic helix at the N-terminus and an aromatic amino acid-rich segment at the site of membrane binding. |
| 巻・号 | 7(2) |
| ページ | 407-22 |
| 公開日 | 2015-2-3 |
| DOI | 10.3390/toxins7020407 |
| PII | toxins7020407 |
| PMID | 25654788 |
| PMC | PMC4344632 |
| MeSH | Amino Acid Sequence Animals Electrophoresis, Polyacrylamide Gel Erythrocytes / drug effects Erythrocytes / pathology HeLa Cells Hemolysis / drug effects* Humans Hydra / metabolism* Marine Toxins / genetics Marine Toxins / isolation & purification Marine Toxins / toxicity* Molecular Sequence Data Multigene Family* Mutagenesis, Site-Directed Pore Forming Cytotoxic Proteins / genetics Pore Forming Cytotoxic Proteins / isolation & purification Pore Forming Cytotoxic Proteins / toxicity* Recombinant Proteins |
| IF | 3.531 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
|
| 各媒体での言及数の合計 | 0 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | HeLa(RCB0007) |