Kinetic analysis of F-actin depolymerization in polymorphonuclear leukocyte lysates indicates that chemoattractant stimulation increases actin filament number without altering the filament length distribution.
Kinetic analysis of F-actin depolymerization in polymorphonuclear leukocyte lysates indicates that chemoattractant stimulation increases actin filament number without altering the filament length distribution.
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DOI：
10.1083/jcb.115.3.677
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发表时间：
1991-11
期刊：
影响因子：
--
通讯作者：
Zigmond SH
中科院分区：
文献类型：
--
作者：
Cano ML;Lauffenburger DA;Zigmond SH
丝状肌动蛋白（F-actin）解聚的速率与去极化丝的数量成正比，速率的变化与丝数的变化成正比。为了确定多形核白细胞中肌动蛋白丝的数量和长度，以及在化学引诱剂刺激下f -肌动蛋白增加过程中丝的数量和长度的变化，我们检测了对照细胞和肽刺激细胞裂解物中f -肌动蛋白解聚的时间过程。用TRITC标记的球状肌动蛋白phalloidin染色定量测定F-actin。在1.2 M KCl和10 μ M DNase I中裂解，f -肌动蛋白结合蛋白和g -肌动蛋白分别对解聚动力学的影响最小。为了从解聚时间过程中确定丝的数量和长度，解聚动力学必须受到肌动蛋白单体解离率的限制。比较细胞松弛素存在（无尖端）或不存在（有刺端和无尖端）时解聚的时间过程表明，解聚发生在丝的两端，单体解离是限速的。对照细胞有1.7 +/- 0.4 × 10(5)条丝，平均长度为0.29 +/- 0.09微米。在室温下以0.02 μ m n -甲酰基去亮基亮基苯丙氨酸刺激90 s，可使f -肌动蛋白增加2倍，肌动蛋白丝总数增加约2倍，达到4.0 +/- 0.5 × 10(5)条，平均长度为0.27 +/- 0.07微米。在这两种情况下，大多数（大约80%）的细丝都很短（小于或等于0.18微米）。受激细胞和对照细胞肌动蛋白丝的长度分布相似。
The rate of filamentous actin (F-actin) depolymerization is proportional to the number of filaments depolarizing and changes in the rate are proportional to changes in filament number. To determine the number and length of actin filaments in polymorphonuclear leukocytes and the change in filament number and length that occurs during the increase in F-actin upon chemoattractant stimulation, the time course of cellular F-actin depolymerization in lysates of control and peptide- stimulated cells was examined. F-actin was quantified by the TRITC- labeled phalloidin staining of pelletable actin. Lysis in 1.2 M KCl and 10 microM DNase I minimized the effects of F-actin binding proteins and G-actin, respectively, on the kinetics of depolymerization. To determine filament number and length from a depolymerization time course, depolymerization kinetics must be limited by the actin monomer dissociation rate. Comparison of time courses of depolymerization in the presence (pointed ends free) or absence (barbed and pointed ends free) of cytochalasin suggested depolymerization occurred from both ends of the filament and that monomer dissociation was rate limiting. Control cells had 1.7 +/- 0.4 x 10(5) filaments with an average length of 0.29 +/- 0.09 microns. Chemo-attractant stimulation for 90 s at room temperature with 0.02 microM N-formylnorleucylleucylphenylalanine caused a twofold increase in F-actin and about a two-fold increase in the total number of actin filaments to 4.0 +/- 0.5 x 10(5) filaments with an average length of 0.27 +/- 0.07 microns. In both cases, most (approximately 80%) of the filaments were quite short (less than or equal to 0.18 micron). The length distributions of actin filaments in stimulated and control cells were similar.
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影响因子：
2.9
作者：
RUBINO, S;SMALL, JV
通讯作者：
SMALL, JV
趋化肽诱导的F-肌动蛋白含量，F-肌动蛋白分布和中性粒细胞形状的动力学的动力学。
影响因子：
7.8
作者：
HOWARD, TH;ORESAJO, CO
通讯作者：
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7.8
作者：
CARSON, M;WEBER, A;ZIGMOND, SH
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影响因子：
7.8
作者：
Schliwa, M;van Blerkom, J
通讯作者：
van Blerkom, J
影响因子：
7.8
作者：
HARTWIG, JH;SHEVLIN, P
通讯作者：
SHEVLIN, P