| Study on the Penetrability of PEP-1-P27mt for Cell Membranes in Vitro |
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| 作者姓名： | 严世荣 朱明磊 邱方城 王立林 屈伸 |
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| 作者单位： | Department of Clinical Laboratory Shiyan People's Hospital Affiliated to Yunyang Medical College，Department of Clinical Laboratory Shiyan People's Hospital Affiliated to Yunyang Medical College，Department of Clinical Laboratory Shiyan People's Hospital Affiliated to Yunyang Medical College，Department of Clinical Laboratory Shiyan People's Hospital Affiliated to Yunyang Medical College，Department of Biochemistry and Molecular Biology School of Basic Medical Sciences Tongji Medical College Huazhong University of Science and Technology，Shiyan 442000 China，Shiyan 442000 China，Shiyan 442000 China，Shiyan 442000 China，Wuhan 430030 China |
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| 基金项目： | a grant from Department of Education of Hubei Province, China (No. Q200524001). |
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| 摘 要： | Double-stranded oligomeric nucleotide encoding PEP-1 peptides was synthesized, pro- karyotic expression pET15b-pep-1-p27mt recombinant constructed, E. coli BL21 (DE3)pLysS trans- formed and induced with IPTG to highly express fusion protein PEP-1-P27mt. Fusion protein with an N-terminal His-tag could be purified by Ni2 -resin affinity chromatography and identified by SDS-PAGE and Western blotting. Cultured EC9706 cells treated with PEP-1-P27mt revealed that PEP-1-P27mt was transduced into cells after 15 min and reached maximal intracellular concentra- tions in 2 h. PEP-1-P27mt of 1 μmol/L final concentration could most strongly suppress the growth. It was suggested that PEP-1 can carry P27mt across membrane, which provides a new biological pro- tocol for using cyclin dependent kinase inhibitors p27mt in suppressing the growth of tumor cells.
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Study on the Penetrability of PEP-1-P27mt for Cell Membranes in Vitro |
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| YAN Shirong,ZHU Minglei,QIU Fangcheng,WANG Lilin,QU Shen.Study on the Penetrability of PEP-1-P27mt for Cell Membranes in Vitro[J].Journal of Zuazhong University of Science and Technology: Medical Edition,2007,27(3). |
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| Authors: | YAN Shirong ZHU Minglei QIU Fangcheng WANG Lilin QU Shen |
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| Abstract: | Double-stranded oligomeric nucleotide encoding PEP-1 peptides was synthesized, prokaryotic expression pET15b-pep-1-p27mt recombinant constructed, E. coli BL21 (DE3)pLysS transformed and induced with IPTG to highly express fusion protein PEP-1-P27mt. Fusion protein with an N-terminal His-tag could be purified by Ni2+-resin affinity chromatography and identified by SDS-PAGE and Western blotting. Cultured EC9706 cells treated with PEP-1-P27mt revealed that PEP-1-P27mt was transduced into cells after 15 min and reached maximal intracellular concentrations in 2 h. PEP-1-P27mt of 1 μmol/L final concentration could most strongly suppress the growth. It was suggested that PEP-1 can carry P27mt across membrane, which provides a new biological protocol for using cyclin dependent kinase inhibitors p27mt in suppressing the growth of tumor cells. |
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| Keywords: | cell-penetrating peptides PEP-1 p27mt pET15b-pep-1-p27mt recombinant |
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