Cloning of cDNAs encoding multidrug-resistance (MDR) associated, P-glycoprotein related protein (mini-P-glycoprotein) overexpressed in murine MDR cells.

编码多药耐药性 (MDR) 相关的 cDNA 的克隆，P-糖蛋白相关蛋白（微型 P-糖蛋白）在小鼠 MDR 细胞中过表达。

• 批准号: 07671193
• 负责人: KAWAI Kazuo
• 金额: $ 1.47万
• 依托单位: Mie University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07671193/
• 关键词: Cancer Chemotherapy; Multidrug Resistance; Multiderug Resistance Associated Protein; P-glycoprotein; 化学療法; 抗癌剤; 遺伝子クローニング; 単クローン抗体

As reported previously, an approximately 65 kDa cellular protein, designated mini-P-glycoprotein (Pgp_<mini>), was originally identified in multidrug resistant (MDR) murine cell lines. In those studies, the Pgp_<mini> was suggested to share both an antigen epitope and some nucleotide sequences with P-glycoprotein (P-gp), and increased expression of this protein was found to be correlated with the level of MDR,although, in contrast to the P-gp, Pgp_<mini> was neither glycosylated nor phosphorylated. In the present study, we also demonstrated that some human MDR cells overexpress Pgp_<mini> as well as P-gp. Furthermore, we constructed a cDNA library from a highly MDR murine cell line into lambdagt10, and using a modified differential hybridization screening, we have identified cDNA clones which encode Pgp_<mini>. After the subcloning into plasmid vectors, we sequenced them. Sequence homology search of this gene shows it to be most closely related to P-gp, particularly mdrla. To clarify the function, works are currently underway to clone hybridomas producing monoclonal antibodies specific to Pgp_<mini> and to transfect with the cDNA into drug-sensitive cells to express it.

据先前报道，最初在多药耐药（MDR）小鼠细胞系中发现了一种约65 kDa的细胞蛋白，称为mini- p -糖蛋白（Pgp_<mini>）。在这些研究中，Pgp_<mini>被认为与p -糖蛋白（P-gp）共享抗原表位和一些核苷酸序列，并且发现该蛋白的表达增加与MDR水平相关，尽管与P-gp相反，Pgp_<mini>既没有糖基化也没有磷酸化。在本研究中，我们还证实了一些人MDR细胞过表达Pgp_<mini>和P-gp。此外，我们从高耐药小鼠细胞系中构建了编码lambdagt10的cDNA文库，并通过改进的差分杂交筛选，鉴定出编码Pgp_<mini>的cDNA克隆。将其亚克隆成质粒载体后，对其进行测序。序列同源性分析表明，该基因与P-gp，尤其是与mdrla的亲缘关系最为密切。为了明确其功能，目前正在进行的工作是克隆产生Pgp_<mini>特异性单克隆抗体的杂交瘤，并将cDNA转染到药物敏感细胞中表达。

项目成果

K.Kawai: "Identification of a Prglycepnotein related protein(mini-P-glycaprsfein)which is ounexpeped in multdray relifant old" Biochemical and Bioplyoical Reseauh Communication. 198. 804-810 (1994)

K.Kawai：“Prglycepnotein 相关蛋白（mini-P-glycaprsfein）的鉴定，该蛋白在多日修复旧中是未曾预料到的”生化和生物胶体研究通讯。