Ben-Ze'ev A, Abulafia R
Mol Cell Biol. 1983 Apr;3(4):684-92. doi: 10.1128/mcb.3.4.684-692.1983.
The association of glycoconjugates with the cytoskeletal framework was examined in detergent-extracted cells. Sparse cultures of fibroblasts that assemble only minimal amounts of extracellular matrix were extracted under mild conditions with Triton X-100 which remove most of the lipids and soluble cellular proteins. The detergent-resistant framework retains lectin binding sites in the nucleus, in the perinuclear area occupied by the rough endoplasmic reticulum-Golgi system of the intact cell, and in a network throughout the cytoskeletal framework. Fluorescent-antibody staining with antibody against collagen type I and fibronectin reveals extensive perinuclear staining of the remnant rough endoplasmic reticulum-Golgi system. In contrast, only sporadic staining of the pericellular area is obtained with these antibodies, in sparse cultures of whole cells. Lectin binding sites were detected in the nucleus and are attributed to chromatin-associated glycoconjugates. They can be removed by DNase under conditions that preserve the cytoplasmic lectin binding sites and the nuclear matrix. The results suggest a high degree of integration of the membrane residues of the cytoplasmic elements and the nuclear matrix with the skeletal framework and indicate a possible role for the glycoconjugates in this structural integration.
在经去污剂处理的细胞中研究了糖缀合物与细胞骨架框架的关联。用仅组装极少量细胞外基质的成纤维细胞稀疏培养物在温和条件下用 Triton X-100 进行处理，该去污剂可去除大部分脂质和可溶性细胞蛋白。抗去污剂框架在细胞核、完整细胞的粗面内质网 - 高尔基体系统占据的核周区域以及贯穿细胞骨架框架的网络中保留凝集素结合位点。用抗 I 型胶原和纤连蛋白的抗体进行荧光抗体染色显示，残留的粗面内质网 - 高尔基体系统有广泛的核周染色。相比之下，在全细胞的稀疏培养物中，用这些抗体仅获得细胞周区域的散在染色。在细胞核中检测到凝集素结合位点，这归因于与染色质相关的糖缀合物。在保留细胞质凝集素结合位点和核基质的条件下，它们可被 DNase 去除。结果表明细胞质成分和核基质的膜残余物与骨架框架高度整合，并表明糖缀合物在这种结构整合中可能发挥作用。
