Fractional attachment of CD47 (IAP) to the erythrocyte cytoskeleton and visual colocalization with Rh protein complexes
Fractional attachment of CD47 (IAP) to the erythrocyte cytoskeleton and visual colocalization with Rh protein complexes
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DOI：
10.1182/blood-2002-04-1187
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发表时间：
2003-02-01
期刊：
影响因子：
20.3
通讯作者：
Discher, DE
中科院分区：
文献类型：
--
作者：
Dahl, KN;Westhoff, CM;Discher, DE
CD 47和RhAG以及Rh蛋白的相互作用在彼此之间以及与完整红细胞的细胞骨架之间可视化。在第一项研究中，CD 47用藻红蛋白（PhE）标记的抗体标记，该抗体产生反映CD 47诱导簇的离散斑点。Rh和RhAG相互共定位，并与这些诱导的集群，而带3和血型糖蛋白C保持更均匀地分散在细胞周边。在第二项研究中，将红细胞吸入微量移液器中，并对CD 47和RhAG进行表面梯度的免疫荧光图确定细胞骨架连接。正常红细胞上的CD 47和RhAG梯度被证明几乎相同，并且似乎也介于流体双层和网络连接血型糖蛋白C的梯度之间。类似的梯度，获得CD 47的Rh-空细胞，表明连接的CD 47的血影蛋白-肌动蛋白骨架是独立的Rh或RhAG，并不受CD 47的减少表面表达这些细胞。结果表明，CD 47与Rh和RhAG共定位，但与这些和其他参与细胞骨架附着的主要膜蛋白无关地部分附着在红细胞膜骨架上。结果表明CD 47的均匀碱基分布，受细胞骨架连接的限制，加上较小部分的CD 47，其能够在膜中扩散。(C)2003年，美国血液学会。
Interactions of CD47 and RhAG and the Rh proteins are visualized between one another and with the cytoskeleton of intact erythrocytes. In a first study, CD47 is labeled with a phycoerythrin (PhE)tagged antibody, which generates discrete spots that reflect induced clusters of CD47. Rh and RhAG colocalize with each other and to these induced clusters, whereas Band 3 and glycophorin C remain more homogeneously dispersed on the cell periphery. In a second study, red cells are aspirated into a micropipette, and immunofluorescent maps of the surface gradients that develop for CD47 and RhAG determine cytoskeletal connectivity. CD47 and RhAG gradients on normal red cells prove to be nearly identical and also appear intermediate to those found for the fluid bilayer and network-linked glycophorin C. Similar gradients are obtained for CD47 on Rh-null cells, suggesting that linkage of CD47 to the spectrin-actin skeleton is independent of Rh or RhAG and is not affected by CD47's reduced surface expression on these cells. The results show that CD47 colocalizes with Rh and RhAG but is fractionally attached to the red cell membrane skeleton independent of these and other major integral membrane proteins involved in cytoskeletal attachment. The results imply a homogeneous base distribution of CD47, restrained by cytoskeleton linkages, plus a smaller fraction of CD47, which is able to diffuse in the membrane. (C) 2003 by The American Society of Hematology.