Cell-penetrating peptide PEP-1-mediated transduction of enhanced green fluorescent protein into human umbilical vein endothelial cells / 中国医学科学院学报
Acta Academiae Medicinae Sinicae
;
(6): 93-97, 2007.
Article
in Chinese
| WPRIM
| ID: wpr-230025
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the penetrating ability of fusion protein PEP-1-EGFP with human umbilical vein endothelial cells.</p><p><b>METHODS</b>Two prokaryotic expression plasmids pET15b-EGFP and pET15b-PEP-1-EGFP were constructed and transformed into E. coli BL21 (DE3) to express EGFP and fusion protein PEP-1-EGFP, respectively. The expressed EGFP and PEP-1-EGFP were purified with Ni(2+) -resin affinity chromatography, and their capabilities of transduction into human umbilical vein endothelial cells were evaluated. The time- and dose-dependent transduction of the fusion protein PEP-1-EGFP and its stability in the human umbilical vein endothelial cells were observed. The toxicity of the fusion protein PEP-1-EGFP was detected by MTT method.</p><p><b>RESULTS</b>EGFP failed to be transduced into human umbilical vein endothelial cells, whereas PEP-1-EGFP fusion protein was transduced into cells shortly in 5 minutes. Its transduction was time- and dose-dependent and the fluorescence in the cells were detected even 27 hours later. No cytotoxicity of the fusion protein PEP-1-EGFP to human umbilical vein endothelial cells was detected even when the dose reached up to 200 micromol/L.</p><p><b>CONCLUSION</b>PEP-1-EGFP fusion protein can efficiently transduce the target protein into human umbilical vein endothelial cells, which provides a basis for future researches on the transduction of antioxidant enzymes mediated by the cell-penetrating peptide, PEP-1, in ischemia-reperfusion injury therapy.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Peptides
/
Umbilical Veins
/
Recombinant Fusion Proteins
/
Cells, Cultured
/
Protein Transport
/
Cysteamine
/
Cell Biology
/
Endothelial Cells
/
Green Fluorescent Proteins
/
Toxicity
Limits:
Humans
Language:
Chinese
Journal:
Acta Academiae Medicinae Sinicae
Year:
2007
Type:
Article
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