A membrane cytoskeleton from Dictyostelium discoideum. II. Integral proteins mediate the binding of plasma membranes to F-actin affinity beads.
A membrane cytoskeleton from Dictyostelium discoideum. II. Integral proteins mediate the binding of plasma membranes to F-actin affinity beads.
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来自盘基网柄菌的膜细胞骨架。
DOI：
10.1083/jcb.99.1.58
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发表时间：
1984
期刊：
影响因子：
--
通讯作者：
Ingalls,HM
中科院分区：
文献类型：
--
作者：
Luna,EJ;Goodloe-Holland,CM;Ingalls,HM
在一种新型的低速沉淀法中，高度纯化、超声处理的盘状盘肌桥状菌质膜片段与f -肌动蛋白珠结合（荧光素标记的f -肌动蛋白在抗荧光素IgG-Sephacryl S-1000珠上）。结合被发现是(a)特异性的，因为含有结合荧光素标记的卵清蛋白的珠粒或没有结合荧光素标记的蛋白的珠粒不能与膜结合，(b)在每微克珠粒结合的f -肌动蛋白约0.6微克时可饱和，(c)快速，1/2为4-20分钟，(d)显然具有合理的亲和力，因为脱落速度太慢，无法用现有技术测量。通过低速沉降实验，我们发现超声质膜碎片，在用超聚物提取后，仍然结合f -肌动蛋白珠。热变性膜、蛋白水解膜和盘状豆脂质囊泡不能结合f -肌动蛋白珠。这些结果表明，完整膜蛋白负责超声膜片段与f -肌动蛋白珠的结合。这一发现与先前的观察一致，即整合蛋白介导了盘状蝶胞质膜与溶液中f -肌动蛋白之间的相互作用（Luna, e.j., v.m. Fowler， J. Swanson， D. Branton, and D. L. Taylor, 1981， J. Cell Biol）。, 88:396 - 409)。我们得出结论，使用f -肌动蛋白珠进行低速沉降试验是监测f -肌动蛋白与膜之间关联的可靠方法。由于这些检测是相对定量的，只需要微克的膜和f -肌动蛋白，它们是对其他现有技术的重大改进，用于探索f -肌动蛋白-膜相互作用的生化细节。使用f -肌动蛋白珠作为肌动蛋白结合蛋白的亲和柱，我们发现至少有12个完整的多肽在盘状豆科植物的质膜上直接或间接地与f -肌动蛋白结合。这些多肽中至少有四种似乎跨越了细胞膜，因此是细胞骨架和细胞表面之间直接跨膜连接的候选者。
In novel, low-speed sedimentation assays, highly purified, sonicated Dictyostelium discoideum plasma membrane fragments bind to F-actin beads (fluorescein-labeled F-actin on antifluorescein IgG-Sephacryl S-1000 beads). Binding was found to be (a) specific, since beads containing bound fluorescein-labeled ovalbumin or beads without bound fluorescein-labeled protein do not bind membranes, (b) saturable at approximately 0.6 microgram of membrane protein per microgram of bead-bound F-actin, (c) rapid with a t1/2 of 4-20 min, and (d) apparently of reasonable affinity since the off rate is too slow to be measured by present techniques. Using low-speed sedimentation assays, we found that sonicated plasma membrane fragments, after extraction with chaotropes, still bind F-actin beads. Heat-denatured membranes, proteolyzed membranes, and D. discoideum lipid vesicles did not bind F-actin beads. These results indicate that integral membrane proteins are responsible for the binding between sonicated membrane fragments and F-actin on beads. This finding agrees with the previous observation that integral proteins mediate interactions between D. discoideum plasma membranes and F-actin in solution (Luna, E.J., V. M. Fowler, J. Swanson, D. Branton, and D. L. Taylor, 1981, J. Cell Biol., 88:396-409). We conclude that low-speed sedimentation assays using F-actin beads are a reliable method for monitoring the associations between F-actin and membranes. Since these assays are relatively quantitative and require only micrograms of membranes and F-actin, they are a significant improvement over other existing techniques for exploring the biochemical details of F-actin-membrane interactions. Using F-actin beads as an affinity column for actin-binding proteins, we show that at least 12 integral polypeptides in D. discoideum plasma membranes bind to F-actin directly or indirectly. At least four of these polypeptides appear to span the membrane and are thus candidates for direct transmembrane links between the cytoskeleton and the cell surface.
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稳定、高容量的 F-肌动蛋白亲和柱。
DOI：
--
发表时间：
1982
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Luna,EJ;Wang,YL;VossJr,EW;Branton,D;Taylor,DL
通讯作者：
Taylor,DL
来自鸡交感神经元和鸡胚脑的细胞骨架相关糖蛋白。
DOI：
10.1111/j.1432-1033.1983.tb07651.x
发表时间：
1983
期刊：
European journal of biochemistry
影响因子：
--
作者：
D. Moss
通讯作者：
D. Moss
肌动蛋白与从盘基网柄菌分离的质膜的细胞质表面结合。
DOI：
10.1016/s0006-291x(80)80034-9
发表时间：
1980
影响因子：
3.1
作者：
Bruce S. Jacobson
通讯作者：
Bruce S. Jacobson
细胞骨架结构对表面受体迁移性的跨膜控制
影响因子：
64.8
作者：
I. Yahara
通讯作者：
I. Yahara
高亲和力兔抗荧光 IgG 抗体亚群与同源配体之间的一级解离率。
影响因子：
3.6
作者：
R. M. Watt;J. Herron;E. Voss
通讯作者：
E. Voss