GTPgammaS-induced actin polymerisation in vitro: ATP- and phosphoinositide-independent signalling via Rho-family proteins and a plasma membrane-associated guanine nucleotide exchange factor.
GTPgammaS-induced actin polymerisation in vitro: ATP- and phosphoinositide-independent signalling via Rho-family proteins and a plasma membrane-associated guanine nucleotide exchange factor.
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GTPgammaS 诱导的体外肌动蛋白聚合：通过 Rho 家族蛋白和质膜相关鸟嘌呤核苷酸交换因子实现 ATP 和磷酸肌醇独立信号传导。
DOI：
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发表时间：
1998
影响因子：
4
通讯作者：
Matthias P. Wymann
中科院分区：
文献类型：
--
作者：
V. Katanaev;Matthias P. Wymann
在中性粒细胞胞浆的无细胞体系中，S可诱导游离细丝带刺末端数目的增加和大量的肌动蛋白聚合和交联。GTP(&ggr；)S的刺激对过量的GDP值敏感，但不能被百日咳波氏杆菌毒素所刺激，并且不能被三聚体G蛋白的氟化铝、肉豆蔻酰化的GTP-Gamma、Gialpha2或Gbeta1-Gamma2亚基所模拟。相反，RhoGDI和艰难梭菌毒素B(灭活Rho家族蛋白)完全取消了GTP GammaS的作用。然而，当以内源性100倍的浓度单独或联合检测重组的、成分激活的和GTPGammaS负载的rac1、RhoA或CDC42蛋白时，它们是无效的。纯化的WASP或C3转移酶的CDC42/RAC相互作用结合(CRIB)结构域不能阻止GTP-GammaS引起的肌动蛋白聚合。GTP-GammaS的作用可被Mm[Mg~(2+)]阻断，除非加入中性粒细胞质膜上存在的热和胰酶敏感成分。因此，带刺末端的释放似乎是由毒素B敏感的胞浆Rho家族蛋白介导的，在生理条件下，需要膜相关的鸟嘌呤核苷酸交换因子(Global)才能被GTP GammaS激活。不同的蛋白激酶和磷酸酶抑制剂(星形孢子素、染料木素、Wortmannin、冈田酸和钒酸)的效率低下，以及apyrase对ATP的去除，表明GTP-GammaS信号的下游传播不需要磷酸转移反应。此外，外源添加的磷脂酰肌醇不能诱导肌动蛋白聚合，PtdIns(4，5)P2结合肽不干扰对GTPGammaS的反应。应用于蛋白质纯化技术的快速和简单的分析方法将有助于进一步阐明参与肌动蛋白聚合的分子伙伴。
In a cell-free system from neutrophil cytosol GTP(&ggr ;)S can induce an increase in the number of free filament barbed ends and massive actin polymerisation and cross-linking. GTP(&ggr ;)S stimulation was susceptible to an excess of GDP, but not Bordetella pertussis toxin and could not be mimicked by aluminium fluoride, myristoylated GTPgammaS.Gialpha2 or Gbeta1gamma2 subunits of trimeric G proteins. In contrast, RhoGDI and Clostridium difficile toxin B (inactivating Rho family proteins) completely abrogated the effect of GTPgammaS. When recombinant, constitutively activated and GTPgammaS-loaded Rac1, RhoA, or Cdc42 proteins alone or in combination were probed at concentrations >100 times the endogenous, however, they were ineffective. Purified Cdc42/Rac-interactive binding (CRIB) domain of WASP or C3 transferase did not prevent actin polymerisation by GTPgammaS. The action of GTPgammaS was blocked by mM [Mg2+], unless a heat- and trypsin-sensitive component present in neutrophil plasma membrane was added. Liberation of barbed ends seems therefore to be mediated by a toxin B-sensitive cytosolic Rho-family protein, requiring a membrane-associated guanine nucleotide exchange factor (GEF) for its activation by GTPgammaS under physiologic conditions. The inefficiency of various protein kinase and phosphatase inhibitors (staurosporine, genistein, wortmannin, okadaic acid and vanadate) and removal of ATP by apyrase, suggests that phosphate transfer reactions are not required for the downstream propagation of the GTPgammaS signal. Moreover, exogenously added phosphoinositides failed to induce actin polymerisation and a PtdIns(4,5)P2-binding peptide did not interfere with the response to GTPgammaS. The speed and simplicity of the presented assay applicable to protein purification techniques will facilitate the further elucidation of the molecular partners involved in actin polymerisation.
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影响因子：
2.9
作者：
HUANG, ZJ;HAUGLAND, RP;HAUGLAND, RP
通讯作者：
HAUGLAND, RP
鸟嘌呤核苷酸交换调节 Rac/Rho GTP 结合蛋白的膜易位。
DOI：
--
发表时间：
1994
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Bokoch,GM;Bohl,BP;Chuang,TH
通讯作者：
Chuang,TH
G 蛋白 α 亚基在大肠杆菌中的表达。
影响因子：
--
作者：
Lee,E;Linder,ME;Gilman,AG
通讯作者：
Gilman,AG
生物活性脂质是 Rac.GDI 络合的调节剂。
DOI：
--
发表时间：
1993
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Chuang,TH;Bohl,BP;Bokoch,GM
通讯作者：
Bokoch,GM
DOI：
10.1016/0006-291x(80)91175-4
发表时间：
1980-01-01
影响因子：
3.1
作者：
MACLEANFLETCHER, S;POLLARD, TD
通讯作者：
POLLARD, TD