Receptor mediated uptake of peptides that bind the human transferrin receptor
Receptor mediated uptake of peptides that bind the human transferrin receptor
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DOI：
10.1046/j.1432-1327.2001.02073.x
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发表时间：
2001-04-01
期刊：
影响因子：
--
通讯作者：
Moore, BA
中科院分区：
文献类型：
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作者：
Lee, JH;Engler, JA;Moore, BA
为了寻找细胞表面受体的特异性多肽表位，本研究采用生物扫描技术，筛选出能与人转铁蛋白受体(HTfR)结合和内化的7个和12个氨基酸的多肽。通过连续几轮的阴性和阳性选择，鉴定出两个与hTfR特异结合的多肽序列。含有HAIYPRH或THRPPMWSPVWP序列的噬菌体在竞争实验中存在相同序列的多肽时，以剂量依赖的方式抑制hTfR的结合。有趣的是，转铁蛋白并不与这两个序列中的任何一个竞争受体结合，这表明这些多肽与hTfR上的一个不同于转铁蛋白结合位点的位点结合。当这些序列中的任何一个被表达为与绿色荧光蛋白(GFP)的融合时，重组GFP分子被内化到表达hTfR的细胞中。这些研究表明，这两种多肽可以用来靶向其他蛋白质进入内体途径。此外，它们提供了一种识别与其他细胞表面受体结合的多肽的策略，这些受体既可用于诊断，也可用于治疗目的。
A biopanning process designed to find peptide epitopes specific for cell surface receptors has been used in this study to select seven- and 12-amino-acid peptides capable of binding to and internalizing with the human transferrin receptor (hTfR). Through sequential rounds of negative and positive selection, two peptide sequences were identified that specifically bind to the hTfR. Phage containing the sequences HAIYPRH or THRPPMWSPVWP were inhibited from binding the hTfR in a dose-dependent fashion when peptides of the same sequence were present in a competition assay. Interestingly, transferrin did not compete with either of these sequences for receptor binding, suggesting that these peptides bind a site on the hTfR distinct from the transferrin binding site. When either of these sequences was expressed as a fusion to green fluorescent protein (GFP), the recombinant GFP molecule was internalized in cells expressing the hTfR. These studies suggest that the two peptides can be used to target other proteins into the endosomal pathway. Further, they provide a strategy for identifying peptides that bind to other cell surface receptors that can be used for both diagnostic and therapeutic purposes.