Coupling between NMDA receptor and acid-sensing ion channel contributes to ischemic neuronal death

Acid-sensing ion channels (ASICs) composed of ASIC1a subunit exhibit a high Ca2+ permeability and play important roles in synaptic plasticity and acid-induced cell death. Here, we show that ischemia enhances ASIC currents through the phosphorylation at Ser478 and Ser479 of ASIC1a, leading to exacerbated ischemic cell death. The phosphorylation is catalyzed by Ca2+/calmodulin-dependent protein kinase II (CaMKII) activity, as a result of activation of NR2B-containing N-methyl-D-aspartate subtype of glutamate receptors (NMDARs) during ischemia. Furthermore, NR2B-specific antagonist, CaMKII inhibitor, or overexpression of mutated form of ASIC1a with Ser478 or Ser479 replaced by alanine (ASICla-S478A, ASIC1a-S479A) in cultured hippocampal neurons prevented ischemia-induced enhancement of ASIC currents, cytoplasmic Ca2+ elevation, as well as neuronal death. Thus, NMDAR-CaMKII cascade is functionally coupled to ASICs and contributes to acidotoxicity during ischemia. Specific blockade of NMDAR/CaMKII-ASIC coupling may reduce neuronal death after ischemia and other pathological conditions involving excessive glutamate release and acidosis.

稀土离子对钙调蛋白及其靶酶结构与功能的影响

稀土元素由于在农业上的广泛应用已经越来越多地进入到生物体循环中，因此在分子水平上研究它们对生物体功能的影响及其作用机理成为人们迫切需要解决的课题。钙调蛋白（CaM）是广泛存在于生物体内的一种多功能的调节蛋白，调节着至少40余种酶的活性。基于上述原因，本文选取钙调蛋白作为研究对象，系统地比较了不同浓度的钙离子和稀土离子对它的靶酶钙调神经磷酸酶、磷脂酶凡、乳酸脱氢酶活性的影响，并采用荧光光谱、同步荧光光谱、紫外差谱、红外光谱等方法研究了它们对钙调蛋白及其靶酶构象变化的影响。发现稀土离子对钙调蛋白靶酶的活性均表现出"低浓度促进，高浓度抑制"的Hormesis效应，由于靶酶分子结构的差异，其最大激活浓度有所区别，钙调蛋白可以缓解稀土离子对靶酶的抑制作用；发现高浓度的稀土离子与钙调蛋白作用后，可以降低钙调蛋白与靶肤／酶分子的亲合性，并抑制其调节靶酶活性功能的发挥；稀土离子与钙调蛋白作用后，增大其α-螺旋含量，减少价折叠结构，对小肤的二级结构影响较小；二乙三氨五乙酸及其衍生物二乙三胺五乙酸－双二甲酞胺和二乙三胺五乙酸一双（异烟麟）可以作为乳酸脱氢酶的竞争性抑制剂抑制其活性，抑制常数分别为3.37、7.41、8.52μmol几，钙调蛋白也可以缓解它们对脱氢酶的抑制作用；首次制备出一株由稀土离子诱导钙调蛋白构象变化后的单克隆抗体2C3，该抗体可以特异性地识别含稀土的钙调蛋白，与Ca－CaM、La-CaM、Eu-CaM以及Yb-CaM之间的解离常数分别为157、26.8、21.8和64.8 nmol／L，该抗体还能够区分钙调蛋白结合不同浓度稀土离子后的构象状态。

金属离子与钙调蛋白相互作用的基质辅助激光解析电离飞行时间质谱研究

Calmodulin is a ubiquitous calcium-binding protein in eukaryote cells and engages in various important biological pathways. In the present study, binding interactions between several metal ions and calmodulin were nvestigated by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS).The results revealed that the specific binding of metal ions with the protein could be detected using MALDI-TOF MS.

INVOLVEMENT OF CA(2+) SIGNALING PATHWAY DURING OOCYTE MATURATION OF THE NORTHERN QUAHOG MERCENARIA MERCENARIA

In many molluses, it has been found that Ca2+ signaling pathway is involved in the resumption of meiotic maturation in oocytes. To better understand the possible role of Ca2+ signaling pathway in regulating meiotic maturation in oocytes of the northern quahog Mercenaria mercenaria, free extracellular Ca2+, A23187 (calcium ionophore), verapamil (calcium channel blocker), and trifluoperazin (calmodulin antagonist) were used to incubate oocytes or serotonin-induced oocytes by pharmacological methods. Results show that extracellular Ca2+ (50 similar to 200 mM) and A23187 (1 similar to 10 mu M) can stimulate the meiotic maturation. In addition, verapamil (1 similar to 100 mu M) and trifluoperazin (10 similar to 1,000 mu M) could inhibit serotonin-induced oocyte maturation. Therefore, Ca2+ is essential for the reinitiation of meiotic maturation in oocytes of the northern quahog. Moreover, an increase i [Ca2+]i can promote meiotic maturation.