Biological effects of SLURP-1 on human keratinocytes
Biological effects of SLURP-1 on human keratinocytes
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DOI：
10.1111/j.0022-202x.2005.23973.x
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发表时间：
2005-12-01
影响因子：
6.5
通讯作者：
Grando, SA
中科院分区：
文献类型：
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作者：
Arredondo, J;Chernyavsky, AI;Grando, SA
在Mal de Meleda的SLURP（分泌哺乳动物Ly-6/尿激酶型纤溶酶原激活物受体相关蛋白）-1的研究中，发现了一种通过尼古丁乙酰胆碱受体（nAChR）调节角化细胞（KC）的新模式。我们克隆了人SLURP-1，制备了重组蛋白和单克隆抗体336H12-1A3，该单克隆抗体可见于天然SLURP-1。SLURP-1连接了KC nAChR的传统配体结合位点，与[3)H]尼古丁-相比，对[3 H]表贝替丁敏感的nAChR表现出更高的亲和力。SLURP-1显著（p < 0.05）提高了caspase 3和caspase 8的活性以及末端脱氧核苷酸转移酶介导的dutp -生物素缺口末端标记阳性细胞的数量。SLURP-1的促凋亡活性超过肿瘤坏死因子- α，提示其参与不同的通路。在一系列实时PCR和细胞内western实验中，SLURP-1显著（p < 0.05）上调转谷氨酰胺酶I型细胞角蛋白10、p21和caspase-3的表达。在激动剂carbachol的存在下，SLURP-1对基因表达的影响被增强，这与SLURP-1在KC nAChR中作为变构激动剂的概念保持一致。因此，SLURP-1诱导的细胞状态变化可能是nachr介导的KC基因表达的影响所致。这些结果表明，SLURP-1在表皮中的生物学作用是通过胆碱能途径对KC功能的生理调节提供微调。
A novel paradigm of keratinocyte (KC) regulation via nicotinic acetylcholine receptors (nAChR) has been discovered in studies of SLURP (secreted mammalian Ly-6/urokinase-type plasminogen activator receptor-related protein)-1 in Mal de Meleda. We cloned human SLURP-1 and produced recombinant protein and the monoclonal antibody 336H12-1A3 that visualized native SLURP-1. SLURP-1 ligated the conventional ligand-binding site of KC nAChR, showing a higher affinity to the [(3)H]nicotine-, compared with the [3 H]epibatidine-sensitive nAChR. SLURP-1 significantly (p < 0.05) increased the activities of caspases 3 and 8, and the number of terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling-positive cells. The pro-apoptotic activity of SLURP-1 exceeded that of tumor necrosis factor-alpha, suggesting the involvement of separate pathways. In a series of real-time PCR and in-cell western experiments, SLURP-1 significantly (p < 0.05) upregulated expression of transglutaminase type I cytokeratin 10, p21, and caspase-3. In the presence of the agonist carbachol, the effects of SLURP-1 on gene expression were augmented, which is in keeping with the notion that SLURP-1 acts as an allosteric agonist at the KC nAChR. Thus, the changes in the cell state induced by SLURP-1 could result from nAChR-mediated effects on the KC gene expression. These results suggest that the biological role of SLURP-1 in the epidermis is to provide fine tuning of the physiologic regulation of KC functions through the cholinergic pathways.