FEBS Letters | |
Regulation of phospholipase D activity in synaptosomes permeabilized with Staphylococcus aureus α‐toxin | |
Böckmann, Ira2  Klein, Jochen2  Valeva, Angela1  von Eichel-Streiber, Christoph1  Kempter, Ulrike2  Sarri, Elisabet2  Weichel, Oksana2  | |
[1] Department of Medical Microbiology and Hygiene, University of Mainz, Obere Zahlbacher Str. 67, D-55101 Mainz, Germany;Department of Pharmacology, University of Mainz, Obere Zahlbacher Str. 67, D-55101 Mainz, Germany | |
关键词: Synaptosome; Phospholipase D; Staphylococcus aureus α-toxin; Calcium/calmodulin-dependent protein kinase II; Rho protein; Clostridium difficile toxin B; BFA; brefeldin A; CaMKII; calcium/calmodulin-dependent kinase II; PEth; phosphatidylethanol; PKC; protein kinase C; PLD; phospholipase D; PtdCho; phosphatidylcholine; TcdB; Clostridium difficile toxin B; | |
DOI : 10.1016/S0014-5793(98)01479-3 | |
学科分类:生物化学/生物物理 | |
来源: John Wiley & Sons Ltd. | |
【 摘 要 】
In order to investigate the regulation of presynaptic phospholipase D (PLD) activity by calcium and G proteins, we established a permeabilization procedure for rat cortical synaptosomes using Staphylococcus aureus α-toxin (30–100 μg/ml). In permeabilized synaptosomes, PLD activity was significantly stimulated when the concentration of free calcium was increased from 0.1 μM to 1 μM. This activation was inhibited in the presence of KN-62 (1 μM), an inhibitor of calcium/calmodulin-dependent kinase II (CaMKII), but not by the protein kinase C inhibitor, Ro 31-8220 (1–10 μM). Synaptosomal PLD activity was also stimulated in the presence of 1 μM GTPγS. When Rho proteins were inhibited by pretreatment of the synaptosomes with Clostridium difficile toxin B (TcdB; 1–10 ng/ml), the effect of GTPγS was significantly reduced; in contrast, brefeldin A (10–100 μM), an inhibitor of ARF activation, was ineffective. Calcium stimulation of PLD was inhibited by TcdB, but GTPγS-dependent activation was insensitive to KN-62. We conclude that synaptosomal PLD is activated in a pathway which sequentially involves CaMKII and Rho proteins.
【 授权许可】
Unknown
【 预 览 】
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