Reference - Detail
| RRC ID | 38220 |
|---|---|
| Author | Yu HH, Nakase I, Pujals S, Hirose H, Tanaka G, Katayama S, Imanishi M, Futaki S. |
| Title | Expressed protein ligation for the preparation of fusion proteins with cell penetrating peptides for endotoxin removal and intracellular delivery. |
| Journal | Biochim Biophys Acta |
| Abstract |
Expressed protein ligation (EPL) is a useful method for the native chemical ligation of proteins with other proteins or peptides. This study assessed the practicability of EPL in the preparation of fusion proteins of enhanced green fluorescent protein (EGFP) with chemically synthesized cell-penetrating peptides (CPPs) for intracellular delivery. Using intein-mediated purification with an affinity chitin-binding tag (IMPACT) system, the thioester of EGFP (EGFP-SR) was prepared. Optimization of the ligation of EGFP-SR with arginine 12-mer (R12) produced the fusion protein in high yield. The EPL procedure also allows the preparation of EGFP-R12 containing a low level of endotoxin (ET), via the satisfactory ET removal of EGFP-SR prior to ligation with the R12 peptide. Fusion proteins of EGFP with R12 and the d-isomer of R12 prepared by EPL showed similar levels of cellular uptake compared to the fusion protein directly expressed in Escherichiacoli. |
| Volume | 1798(12) |
| Pages | 2249-57 |
| Published | 2010-12-1 |
| DOI | 10.1016/j.bbamem.2010.02.003 |
| PII | S0005-2736(10)00043-X |
| PMID | 20170629 |
| MeSH | Animals Cell-Penetrating Peptides / biosynthesis Cell-Penetrating Peptides / chemistry Cell-Penetrating Peptides / genetics Cell-Penetrating Peptides / isolation & purification Cell-Penetrating Peptides / pharmacology Chromatography, Affinity / methods Endotoxins* Escherichia coli Gene Expression Green Fluorescent Proteins / biosynthesis Green Fluorescent Proteins / chemistry Green Fluorescent Proteins / genetics Green Fluorescent Proteins / pharmacology HeLa Cells Humans Inteins Recombinant Fusion Proteins* / biosynthesis Recombinant Fusion Proteins* / chemistry Recombinant Fusion Proteins* / genetics Recombinant Fusion Proteins* / isolation & purification Recombinant Fusion Proteins* / pharmacology |
| IF | 3.411 |
| Times Cited | 11 |
| WOS Category | BIOPHYSICS BIOCHEMISTRY & MOLECULAR BIOLOGY |
| Altmetric score |
オルトメトリクス指標項目
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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) |