Boonen G J, VanSteveninck J, Elferink J G
Department of Medical Biochemistry, University of Leiden, Netherlands.
Biochim Biophys Acta. 1993 Jan 17;1175(2):155-60. doi: 10.1016/0167-4889(93)90018-k.
Calcium, strontium and barium induced an exocytotic response in electropermeabilized rabbit neutrophils while magnesium was without any effect. The extent of enzyme release was found to depend upon the concentration of these cations. For all cations, an optimum concentration was found with the same maximum enzyme release. At concentrations higher than optimum a decrease in lysozyme release was observed. Efficiency to induce enzyme release was in the order: Ca2+ > Sr2+ > Ba2+. Enzyme release was significantly enhanced by guanosine-5'-[gamma-thio]triphosphate (GTP gamma S) resulting in a shift to the left of the dose/response curve. The enhancement by GTP gamma S was strongest with Ca2+, was less with Sr2+, and was very little with Ba2+. The time course of lysozyme release was the same for Ca2+, Sr2+, and Ba2+ in the presence and absence of GTP gamma S when suboptimal cation concentrations were used. A decrease in responsiveness to the effectors after electropermeabilization was observed with Ca2+, Sr2+ and Ba2+ in the presence and absence of GTP gamma S. The lysozyme release induced by the different cations was not inhibited by the protein kinase C inhibitor staurosporine and was slightly affected by pertussis toxin. Ca2+ and Sr2+, but not Ba2+, potentiated formyl-methionyl-leucyl-phenylalanine (fMet-Leu-Phe) induced enzyme release in intact neutrophils. The divalent cation ionophore A23187 induced enzyme release in the presence of Ca2+ and Sr2+ but not in the presence of Ba2+. The results obtained with electropermeabilized neutrophils indicate that Sr2+ and Ba2+ can act as substitutes for Ca2+ in activating exocytosis, and that permeabilized neutrophils provide the best tool to investigate the effects of alkaline earth ions in exocytosis.
钙、锶和钡可诱导电穿孔处理的兔中性粒细胞发生胞吐反应,而镁则无任何作用。发现酶释放的程度取决于这些阳离子的浓度。对于所有阳离子,都发现了一个最佳浓度,此时酶释放量达到最大值。在高于最佳浓度时,观察到溶菌酶释放量减少。诱导酶释放的效率顺序为:Ca2+ > Sr2+ > Ba2+。鸟苷 - 5'-[γ-硫代]三磷酸(GTPγS)可显著增强酶释放,导致剂量/反应曲线向左移动。GTPγS对Ca2+的增强作用最强,对Sr2+的增强作用较小,对Ba2+的增强作用非常小。当使用次优阳离子浓度时,在有和没有GTPγS的情况下,Ca2+、Sr2+和Ba2+诱导的溶菌酶释放时间进程相同。在有和没有GTPγS的情况下,Ca2+、Sr2+和Ba2+电穿孔处理后对效应物的反应性均降低。不同阳离子诱导的溶菌酶释放不受蛋白激酶C抑制剂星形孢菌素的抑制,且受百日咳毒素的影响较小。Ca2+和Sr2+可增强甲酰 - 甲硫氨酰 - 亮氨酰 - 苯丙氨酸(fMet-Leu-Phe)诱导的完整中性粒细胞的酶释放,但Ba2+无此作用。二价阳离子载体A23187在有Ca2+和Sr2+存在时可诱导酶释放,但在有Ba2+存在时则不能。用电穿孔处理的中性粒细胞获得的结果表明,Sr2+和Ba2+可在激活胞吐作用中替代Ca2+,并且电穿孔处理的中性粒细胞为研究碱土金属离子在胞吐作用中的影响提供了最佳工具。