Cell-penetrating Peptide PEP-1-mediated Transduction of Enhanced Green Fluorescent Protein into Human Umbilical Vein Endothelial Cells

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Acta Academiae Medicinae Sinicae ›› 2007, Vol. 29 ›› Issue (1) : 93-97.

DONG Xiao, WANG Jia-ning, HUANG Yong-zhang, GUO Ling-yun,KONG Xia

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ABSTRACT:Objective To investigate the penetrating ability of fusion protein PEP-1-EGFP with human umbilical vein endothelial cells. Methods Two prokaryotic expression plasmids pET15b-EGFP and pET15b-PEP-1-EGFP were constructed and transformed into E.coli BL21(DE3) to express EGFP and fusion protein PEP-1-EGFP, respectively. The expressed EGFP and PEP-1-EGFP were purified with Ni2+-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. Results 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 μmol/L. Conclusion 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.

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cell-penetratinc peptide / protein transduction / endanced creen fluorescent protein / duman umbillical vein endotdelial cell / iscdemia-reperfusion injury

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DONG Xiao, WANG Jia-ning, HUANG Yong-zhang, GUO Ling-yun,KONG Xia. Cell-penetrating Peptide PEP-1-mediated Transduction of Enhanced Green Fluorescent Protein into Human Umbilical Vein Endothelial Cells. Acta Academiae Medicinae Sinicae. 2007, 29(1): 93-97

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Received	Revised	Published
2006-05-15	1900-01-01	2007-02-28
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2007-02-28

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