Creatine kinase-mediated ATP supply fuels actin-based events in phagocytosis.
Creatine kinase-mediated ATP supply fuels actin-based events in phagocytosis.
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DOI：
10.1371/journal.pbio.0060051
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发表时间：
2008-03-11
期刊：
影响因子：
9.8
通讯作者：
Wieringa B
中科院分区：
文献类型：
--
作者：
Kuiper JW;Pluk H;Oerlemans F;van Leeuwen FN;de Lange F;Fransen J;Wieringa B
吞噬作用需要由肌动蛋白聚合和肌球蛋白运动活性驱动的局部协调的细胞骨架重排。这种肌动球蛋白动力学如何依赖于在吞噬体微区提供ATP通路的系统尚未确定。我们分析了脑型肌酸激酶（CK-B）的作用，CK-B是一种参与高能磷酰转移的酶。我们证明，内源性CK-B在巨噬细胞动员从胞质池和cocumulates与F-肌动蛋白在新生的吞噬体。用XFP标记的CK-B和β-肌动蛋白的活细胞成像揭示了这种分配过程的短暂和特异性。催化性死亡CK-B或CK特异性环肌酸抑制的过表达导致吞噬杯区肌动蛋白蓄积显著减少，并降低补体受体介导的巨噬细胞摄取能力，但不降低Fc-γ R介导的巨噬细胞摄取能力。最后，我们发现CK-B的抑制已经在颗粒粘附阶段影响吞噬作用，最有可能是通过影响肌动蛋白聚合行为。我们认为巨噬细胞中的CK-B活性通过提供局部ATP供应而有助于补体诱导的F-actin在早期吞噬作用中的组装事件。为了工作，细胞需要ATP形式的能量。在细胞形状改变过程中，可以看到高而突然的能量需求，这是一个ATP为细胞骨架机制提供燃料的过程，该机制驱动细胞形态改变。细胞如何在不破坏整体ATP稳态的情况下组织高能量激增仍然是一个重要的研究问题。一种观点认为，ATP是不均匀分布的，但细胞骨架蛋白肌动蛋白和肌球蛋白接受区域和优先访问ATP。然而，这个模型提出了另一个问题：ATP是如何从遥远的来源输送到这些蛋白质的？为了解决这些问题中的一些，我们研究了高度本地化的分子事件控制周围的巨噬细胞的吞噬活性肌动蛋白动力学。我们证明，肌动蛋白和肌酸激酶-B（CK-B），一个长期已知的酶参与ATP供应，同时招募到行动的网站在颗粒摄入的早期阶段。CK活性的局部可用性和ATP的局部产生促进了现场肌动蛋白重塑和颗粒捕获效率，从而支持吞噬作用的第一阶段的成功启动。有趣的是，局部CK活性和肌动蛋白调节之间的这种偶联仅与补体介导的吞噬作用有关（免疫细胞用于靶向特定颗粒以供摄取）。我们预测，我们的研究结果也可能揭示形状动力学如何在其他细胞类型中被激活。在巨噬细胞吞噬杯中，肌酸激酶-B（CK-B）的局部募集与肌动蛋白重塑活性之间存在紧密联系。补体介导的吞噬作用在酶活性CK-B的存在下被刺激，表明局部ATP供应刺激肌动蛋白驱动的颗粒摄取事件。
Phagocytosis requires locally coordinated cytoskeletal rearrangements driven by actin polymerization and myosin motor activity. How this actomyosin dynamics is dependent upon systems that provide access to ATP at phagosome microdomains has not been determined. We analyzed the role of brain-type creatine kinase (CK-B), an enzyme involved in high-energy phosphoryl transfer. We demonstrate that endogenous CK-B in macrophages is mobilized from the cytosolic pool and coaccumulates with F-actin at nascent phagosomes. Live cell imaging with XFP-tagged CK-B and β-actin revealed the transient and specific nature of this partitioning process. Overexpression of a catalytic dead CK-B or CK-specific cyclocreatine inhibition caused a significant reduction of actin accumulation in the phagocytic cup area, and reduced complement receptor–mediated, but not Fc-γR–mediated, ingestion capacity of macrophages. Finally, we found that inhibition of CK-B affected phagocytosis already at the stage of particle adhesion, most likely via effects on actin polymerization behavior. We propose that CK-B activity in macrophages contributes to complement-induced F-actin assembly events in early phagocytosis by providing local ATP supply. To do work, cells need energy in the form of ATP. High and sudden energy demand is seen during cell-shape change, a process in which ATP fuels the cytoskeletal machinery that drives cell-morphology alteration. How a cell organizes high-energy surges without disrupting global ATP homeostasis remains an important research question. One view proposes that ATP is heterogeneously distributed, but the cytoskeletal proteins actin and myosin receive regional and preferential access to ATP. Yet this model raises another question: how is ATP funneled to these proteins from distant sources? To address some of these questions, we studied the highly localized molecular events controlling actin dynamics around phagocytic activity of macrophages. We demonstrate that actin and creatine kinase-B (CK-B), a long-known enzyme involved in ATP supply, are simultaneously recruited into the sites of action during the early phases of particle ingestion. Local availability of CK-activity and local generation of ATP promotes on-site actin remodeling and particle capture efficiency, and thus supports successful initiation of the first phases of phagocytosis. Interestingly, this coupling between local CK-activity and actin regulation is only relevant for complement-mediated phagocytosis (used by immune cells to target specific particles for ingestion). We predict that our findings may also shed light on how shape dynamics is energized in other cell types. A tight connection exists between local recruitment of creatine kinase-B (CK-B) and actin remodeling activity in phagocytic cups of macrophages. Complement-mediated phagocytosis is stimulated in the presence of enzymatic-active CK-B, indicating that local ATP supply stimulates actin-driven particle uptake events.
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DOI：
10.1073/pnas.261419298
发表时间：
2001-12-18
影响因子：
11.1
作者：
Dayel, MJ;Holleran, EA;Mullins, RD
通讯作者：
Mullins, RD
巨噬细胞中涉及补体和FC受体介导的吞噬作用的不同细胞骨架结构的分子定义。
DOI：
10.1084/jem.184.2.627
发表时间：
1996-08-01
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Allen LA;Aderem A
通讯作者：
Aderem A
影响因子：
5.5
作者：
HASSAN, NF;RIFAT, S;DOUGLAS, SD
通讯作者：
DOUGLAS, SD
影响因子：
21.3
作者：
Cox, D;Berg, JS;Greenberg, S
通讯作者：
Greenberg, S
影响因子：
3.3
作者：
Hoppe, AD;Swanson, JA
通讯作者：
Swanson, JA