Sai Y, Tamai I, Sumikawa H, Hayashi K, Nakanishi T, Amano O, Numata M, Iseki S, Tsuji A
Department of Pharmaceutics, Faculty of Pharmaceutical Sciences, Kanazawa University, Ishikawa, Japan.
FEBS Lett. 1996 Aug 19;392(1):25-9. doi: 10.1016/0014-5793(96)00778-8.
A polyclonal antibody (anti-PepT1/C) was raised against the rabbit intestinal H(+)-coupled oligopeptide transporter, PepT1. Anti-PepT1/C detected 70-80-kDa protein in crude membranes obtained from rabbit duodenum, jejunum and ileum. PepT1 was localized in the brush-border of the absorptive epithelial cells by subcellular fractionation of membranes on a sucrose density gradient and by immunohistochemistry using light and electron microscopy. Transport activity for cephalosporins and dipeptide expressed in Xenopus laevis oocytes injected with total mRNA obtained from rabbit small intestine was eliminated completely by prehybridization of the mRNA with antisense oligonucleotide against the 5'-coding region of rabbit PepT1 cDNA.
制备了一种针对兔肠道H⁺偶联寡肽转运体PepT1的多克隆抗体（抗-PepT1/C）。抗-PepT1/C在从兔十二指肠、空肠和回肠获得的粗制膜中检测到70 - 80 kDa的蛋白质。通过在蔗糖密度梯度上对膜进行亚细胞分级分离以及使用光学显微镜和电子显微镜进行免疫组织化学，将PepT1定位在吸收性上皮细胞的刷状缘。用从兔小肠获得的总mRNA注射非洲爪蟾卵母细胞中所表达的头孢菌素和二肽的转运活性，通过将mRNA与针对兔PepT1 cDNA 5'-编码区的反义寡核苷酸进行预杂交而完全消除。
