Clustering and immobilization of acetylcholine receptors by the 43-kD protein: a possible role for dystrophin-related protein.
Clustering and immobilization of acetylcholine receptors by the 43-kD protein: a possible role for dystrophin-related protein.
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DOI：
10.1083/jcb.123.3.729
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发表时间：
1993-11
影响因子：
7.8
通讯作者：
Merlie, J P
中科院分区：
文献类型：
--
作者：
Phillips, W D;Noakes, P G;Roberds, S L;Campbell, K P;Merlie, J P
当43-kD突触后蛋白（43 k）在相同细胞中共表达时，在培养的成纤维细胞表面上表达的重组乙酰胆碱受体（AChR）被组织成离散的膜结构域（Froehner，S.C.，C. W. Luetje，P. B. Scotland和J.帕特里克，1990年。Neuron. 5：403-410;菲利普斯，W. D、M. C.科普塔，P.布朗特，警察局。加德纳，J.H.施泰因巴赫和J. P.梅利。1991.科学（Wash）DC）。251：568-570）。在这里，我们表明，AChRs目前的成纤维细胞表面上的转染前43 k被招募到43 k丰富的膜结构域。聚集的乙酰胆碱受体表现出增加的电阻提取与Triton X-100，这表明43 k依赖的连接到细胞骨架。小鼠细胞系C2的肌管自发地显示偶尔AChR/43 k丰富的膜结构域，其直径范围高达15微米，但当43 k在43 k cDNA转染后过表达时表达更多。然而，由重组43 k诱导的膜结构域主要是小的（≤ 2微米）。然后，我们感兴趣的是细胞骨架成分，肌营养不良蛋白相关蛋白（DRP; Tinsley，J.M.，D. J. Blake，A.罗氏大学Fairbrother，J. Riss，B. C. Byth，A. E.作者声明：J. K. Suthers，D. R.爱你的Y H. Edwards和K. E.戴维斯，1992年。自然（伦敦）。360：591-593）促进了乙酰胆碱受体集群的发展。免疫荧光抗DRP染色存在于小鼠胚胎神经肌肉突触AChR聚类的最早阶段，也集中在未转染的C2肌管上的大AChR丰富的结构域。令人惊讶的是，抗DRP染色主要集中在大的，但不是小的AChR集群C2肌管表明，DRP可能主要参与允许AChR集群的增长。
Recombinant acetylcholine receptors (AChRs) expressed on the surface of cultured fibroblasts become organized into discrete membrane domains when the 43-kD postsynaptic protein (43k) is co-expressed in the same cells (Froehner, S.C., C. W. Luetje, P. B. Scotland, and J. Patrick, 1990. Neuron. 5:403-410; Phillips, W. D., M. C. Kopta, P. Blount, P. D. Gardner, J. H. Steinbach, and J. P. Merlie. 1991. Science (Wash. DC). 251:568-570). Here we show that AChRs present on the fibroblast cell surface prior to transfection of 43k are recruited into 43k-rich membrane domains. Aggregated AChRs show increased resistance to extraction with Triton X-100, suggesting a 43k-dependent linkage to the cytoskeleton. Myotubes of the mouse cell line C2 spontaneously display occasional AChR/43k-rich membrane domains that ranged in diameter up to 15 microns, but expressed many more when 43k was overexpressed following transfection of 43k cDNA. However, the membrane domains induced by recombinant 43k were predominantly small (< or = 2 microns). We were then interested in whether the cytoskeletal component, dystrophin related protein (DRP; Tinsley, J. M., D. J. Blake, A. Roche, U. Fairbrother, J. Riss, B. C. Byth, A. E. Knight, J. Kendrick-Jones, G. K. Suthers, D. R. Love, Y. H. Edwards, and K. E. Davis, 1992. Nature (Lond.). 360:591-593) contributed to the development of AChR clusters. Immunofluorescent anti-DRP staining was present at the earliest stages of AChR clustering at the neuromuscular synapse in mouse embryos and was also concentrated at the large AChR-rich domains on nontransfected C2 myotubes. Surprisingly, anti-DRP staining was concentrated mainly at the large, but not the small AChR clusters on C2 myotubes suggesting that DRP may be principally involved in permitting the growth of AChR clusters.