Cryptdin 3 forms anion selective channels in cytoplasmic membranes of human embryonic kidney cells.
Cryptdin 3 forms anion selective channels in cytoplasmic membranes of human embryonic kidney cells.
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DOI：
10.1152/ajpgi.00152.2001
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发表时间：
2002-05
期刊：
影响因子：
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通讯作者：
G. Yue;D. Merlin;M. Selsted;W. Lencer;J. Madara;D. Eaton
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文献类型：
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作者：
G. Yue;D. Merlin;M. Selsted;W. Lencer;J. Madara;D. Eaton
Cryptdins是由位于肠隐窝底部的潘氏细胞分泌的抗微生物肽。除了它们的抗微生物功能之外，Cryptdins还可以调节肠上皮细胞的盐和水分泌。最近的工作与短路电流测量表明，至少有一个cryptdin肽，cryptdin 3，诱导顶端电导在Cl（-）分泌，包括囊性纤维化，上皮细胞。在本研究中，我们的特点cryptdin 3诱导的阴离子通道活动在人胚肾（HEK）细胞与单通道膜片钳技术。贴片移液管充满含有不同浓度的cryptdin 3的溶液，并且在gigaseal形成后，用细胞附着或由内而外的贴片模式记录通道活性。我们发现了一个阴离子选择性通道的电导为15 pS和开放概率为0.19，无论cryptdin 3浓度。平均开放和关闭时间随穴状蛋白3浓度而变化。对于移液管中10、4、1和0.5 μ g/ml的穴状蛋白3浓度，相应的平均开放时间为1.2、7.0、9.0和17.4 ms，相应的平均关闭时间为1.1、1.6、4.2、这些结果表明，cryptdin 3形成阴离子-本发明还涉及HEK细胞的细胞质膜上的选择性通道，并且一个这样的通道的动力学受其与其他这样的通道的相互作用的影响。
Cryptdins are antimicrobial peptides secreted by Paneth cells located at the base of intestinal crypts. In addition to their antimicrobial function, cryptdins may also regulate salt and water secretion by intestinal epithelial cells. Recent work with short-circuit current measurements indicated that at least one cryptdin peptide, cryptdin 3, induces apical conductance(s) in Cl(-) secretory, including cystic fibrosis, epithelia. In the present study, we characterized the cryptdin 3-induced anion channel activity in human embryonic kidney (HEK) cells with single-channel patch-clamp techniques. The patch pipette was filled with solution containing different concentrations of cryptdin 3, and, after gigaseal formation, the channel activity was recorded with either cell-attached or inside-out patch modes. We found an anion selective channel with a conductance of 15 pS and open probability of 0.19, regardless of cryptdin 3 concentration. The mean open and closed times varied with the cryptdin 3 concentration. For cryptdin 3 concentrations of 10, 4, 1, and 0.5 microg/ml in the pipette, the corresponding mean open times were 1.2, 7.0, 9.0, and 17.4 ms and the corresponding mean closed times were 1.1, 1.6, 4.2, and 12.5 ms. These results suggest that cryptdin 3 forms anion-selective channels on the cytoplasmic membrane of HEK cells and that the kinetics of one such channel are affected by its interaction with other such channels.