Expression of P-glycoprotein and Biospanning of Its Specific Peptide
WANG Xiang wei 1) , YANG Tang jun 1) , ZHANG Li xin 2) ,GUO Ying lu 3)
( 1) Department of Urology, The Second affiliated Hospital of The Third Military Medical University, Chongqing 400037,China; 2) Department of Tumor Molecular Biology, Academy of Military Medical Sciences, Beijing 100750,China; 3) Institute of Urology, Peking University, Beijing 100034,China
Abstract：In order to express extracellular fragment of P glycoprotein, a highly expressed recombinant pGEX Pgp was constructed, transferred into DH5α and expressed. The fused protein was identified and purified. The fused P glycoprotein was used as the target to biospan its binding peptide in phage random 12 peptide library, and immunocyto chemical staining in BIU 87/ADM and BIU 87 cells was done to identify the positive phage clones. SDS PAGE analysis indicated that the expressed protein was about 30 kD. Peptide specific binding to P glycoprotein was acquired after 4 runs of biospanning. The inserted sequence of the peptide as NDGLLFTWQPSP was identified by sequencing. Immunocyto chemical staining revealed that the binding occurred only in MDR cells, not in sensitive cells. It indicated that this kind of binding showed its affinity and specificity to MDR cells. The screening of peptide provides a base for the targeted reversal of multidrug resistance in human bladder cancer.
王祥卫,杨唐俊,张立新,郭应禄. P-糖蛋白的表达及其结合肽的筛选[J]. 中国生物化学与分子生物学报, 2003, 19(05): 640-645.
WANG Xiang wei 1) , YANG Tang jun 1) , ZHANG Li xin 2) ,GUO Ying lu 3). Expression of P-glycoprotein and Biospanning of Its Specific Peptide. Chinese Journal of Biochemistry and Molecular Biol, 2003, 19(05): 640-645.
