A D-peptide analog of the second extracellular loop of claudin-3 and -4 leads to mislocalized claudin and cellular apoptosis in mammary epithelial cells.
A D-peptide analog of the second extracellular loop of claudin-3 and -4 leads to mislocalized claudin and cellular apoptosis in mammary epithelial cells.
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DOI：
10.1111/j.1747-0285.2010.01061.x
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发表时间：
2011-02
影响因子：
3
通讯作者：
Neville MC
中科院分区：
文献类型：
--
作者：
Baumgartner HK;Beeman N;Hodges RS;Neville MC
紧密连接蛋白是细胞粘附蛋白，被认为介导紧密连接处的细胞与细胞接触。尽管紧密蛋白的主要作用是控制细胞旁扩散，但越来越多的证据表明它们也可能在肿瘤进展中发挥作用。为了检查第二个细胞外环在细胞粘附中的作用，设计了一种小肽来模拟特定“经典”密蛋白亚型中的保守序列 DFYNP。使用乳腺上皮细胞的荧光指示剂，用该肽的 L 型和 D 型处理显示 Claudin-4 和 Claudin-3 的错误定位以及 caspase-8 和 caspase-3 的激活，表明细胞凋亡。为了测试特异性，制备了具有不同端基并在五个残基中的每个残基处进行甘氨酸取代的肽。改变端基并不影响肽的活性。然而，F147、Y148、N149 或 P150 处的氨基酸取代会阻止肽活性。荧光标记的肽在 4°C 下与紧密连接结合，当培养物升温至 37°C 时会引起细胞凋亡。总之，模拟紧密蛋白第二个细胞外环中的序列的小肽的 D 和 L 形式都可以靶向并破坏上皮单层中的紧密蛋白并引发细胞凋亡。
Claudins are cell adhesion proteins thought to mediate cell-cell contacts at the tight junction. Although a major role of claudins is to control paracellular diffusion, increasing evidence suggests that they may also function in tumor progression. To examine the role of the second extracellular loop in cell adhesion, a small peptide was designed that mimics a conserved sequence, DFYNP, within specific “classic” claudin subtypes. Using fluorescent indicators with mammary epithelial cells, treatment with both the L and D forms of this peptide showed mis-localization of claudin-4 and claudin-3 and activation of caspases-8 and caspase-3, indicating apoptosis. To test specificity, peptides were made both with various end-groups and with glycine substitutions at each of the five residues. Changing end groups did not influence the activity of the peptide. Amino acid substitutions at F147, Y148, N149, or P150, however, prevented peptide activity. A fluorescent labeled peptide was shown to associate with the tight junction at 4°C and cause apoptosis when the cultures were warmed to 37°C. In conclusion, both the D and L forms of a small peptide that mimics a sequence in the second extracellular loop of claudins can target and disrupt claudin proteins in an epithelial monolayer and initiate apoptosis.
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影响因子：
3.1
作者：
Chen, Ming L.;Ge, Zhongming;Schauer, David B.
通讯作者：
Schauer, David B.
影响因子：
5.8
作者：
Hong, SY;Oh, JE;Lee, KH
通讯作者：
Lee, KH
DOI：
10.1152/ajplung.00182.2003
发表时间：
2003-11-01
影响因子：
4.9
作者：
Coyne, CB;Gambling, TM;Johnson, LG
通讯作者：
Johnson, LG
影响因子：
8
作者：
Blasig, IE;Winkler, L;Piontek, J
通讯作者：
Piontek, J
影响因子：
4.1
作者：
Chen, Yuxin;Mant, Colin T.;Hodges, Robert S.
通讯作者：
Hodges, Robert S.