Two chromogranin a-derived peptides induce calcium entry in human neutrophils by calmodulin-regulated calcium independent phospholipase A2.
Two chromogranin a-derived peptides induce calcium entry in human neutrophils by calmodulin-regulated calcium independent phospholipase A2.
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DOI：
10.1371/journal.pone.0004501
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发表时间：
2009
期刊：
影响因子：
3.7
通讯作者：
Metz-Boutigue MH
中科院分区：
文献类型：
--
作者：
Zhang D;Shooshtarizadeh P;Laventie BJ;Colin DA;Chich JF;Vidic J;de Barry J;Chasserot-Golaz S;Delalande F;Van Dorsselaer A;Schneider F;Helle K;Aunis D;Prévost G;Metz-Boutigue MH
来源于嗜铬粒蛋白A（CgA）的天然加工的抗微生物肽在刺激嗜铬细胞时与儿茶酚胺共分泌。由于中性粒细胞在先天免疫中起着核心作用，我们研究了两种抗微生物CgA衍生肽刺激后中性粒细胞的反应。在几种药物的存在下，用不同浓度的CgA衍生肽处理PMN。采用流式细胞术和钙离子成像实验观察钙离子动员情况。免疫细胞化学和共聚焦显微镜显示的肽的细胞内定位。通过表面等离子体共振和iPLA 2活性测定显示肽的钙调蛋白结合和iPLA 2活化性质。最后，通过使用HPLC和Nano-LC MS-MS对用CgA衍生肽处理后的PMNs释放的材料进行蛋白质组学分析。通过使用流式细胞术，我们首先观察到，在15 s后，在细胞外钙的存在下，色芬净（Chromofungin）或Catestatin（CAT）诱导细胞内钙的浓度依赖性瞬时增加。相反，在不存在细胞外钙的情况下，肽在暴露10分钟后不能诱导钙从储存中耗尽。用2-APB（2-氨基乙氧基二苯基硼酸盐）（一种储存操纵通道（SOC）阻断剂）治疗，可完全抑制钙内流，如钙成像所示。我们还表明，它们激活iPLA 2作为两个钙调素结合因子（W7和CMZ），这两个序列可以与iPLA 2报告的两个钙调素结合结构域对齐。最后，我们通过HPLC和Nano-LC MS-MS分析了由PMNs释放的物质，这些物质是由BMPs和CAT刺激后释放的。我们表征了炎症和先天免疫的几个重要因素。这是第一次，我们证明，β-淀粉样蛋白和CAT，渗透到中性粒细胞，诱导细胞外钙离子进入钙调节iPLA 2途径。我们的研究强调了两个CgA衍生肽在神经内分泌和免疫系统之间的积极沟通中的作用。
Antimicrobial peptides derived from the natural processing of chromogranin A (CgA) are co-secreted with catecholamines upon stimulation of chromaffin cells. Since PMNs play a central role in innate immunity, we examine responses by PMNs following stimulation by two antimicrobial CgA-derived peptides. PMNs were treated with different concentrations of CgA-derived peptides in presence of several drugs. Calcium mobilization was observed by using flow cytometry and calcium imaging experiments. Immunocytochemistry and confocal microscopy have shown the intracellular localization of the peptides. The calmodulin-binding and iPLA2 activating properties of the peptides were shown by Surface Plasmon Resonance and iPLA2 activity assays. Finally, a proteomic analysis of the material released after PMNs treatment with CgA-derived peptides was performed by using HPLC and Nano-LC MS-MS. By using flow cytometry we first observed that after 15 s, in presence of extracellular calcium, Chromofungin (CHR) or Catestatin (CAT) induce a concentration-dependent transient increase of intracellular calcium. In contrast, in absence of extra cellular calcium the peptides are unable to induce calcium depletion from the stores after 10 minutes exposure. Treatment with 2-APB (2-aminoethoxydiphenyl borate), a store operated channels (SOCs) blocker, inhibits completely the calcium entry, as shown by calcium imaging. We also showed that they activate iPLA2 as the two CaM-binding factors (W7 and CMZ) and that the two sequences can be aligned with the two CaM-binding domains reported for iPLA2. We finally analyzed by HPLC and Nano-LC MS-MS the material released by PMNs following stimulation by CHR and CAT. We characterized several factors important for inflammation and innate immunity. For the first time, we demonstrate that CHR and CAT, penetrate into PMNs, inducing extracellular calcium entry by a CaM-regulated iPLA2 pathway. Our study highlights the role of two CgA-derived peptides in the active communication between neuroendocrine and immune systems.
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影响因子：
4.8
作者：
Ferrero, E;Scabini, S;Corti, A
通讯作者：
Corti, A
影响因子：
4.8
作者：
Gasparri, A;Sidoli, A;Corti, A
通讯作者：
Corti, A
影响因子：
4.4
作者：
De la Rosa, Gonzalo;Yang, De;Oppenheim, Joost J.
通讯作者：
Oppenheim, Joost J.
影响因子：
6
作者：
Bu, De-xiu;Hemdahl, Anne-Louise;Yan, Zhong-qun
通讯作者：
Yan, Zhong-qun
DOI：
10.1111/j.1432-1033.1996.t01-1-00516.x
发表时间：
1996-02-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
Goumon, Y;Strub, JM;MetzBoutigue, MH
通讯作者：
MetzBoutigue, MH