Monoclonal antibodies distinguish titins from heart and skeletal muscle.
Monoclonal antibodies distinguish titins from heart and skeletal muscle.
复制标题
单克隆抗体将滴虫与心脏和骨骼肌区分开。
DOI：
10.1083/jcb.102.3.1099
复制
发表时间：
1986-03
期刊：
影响因子：
--
通讯作者：
Weber K
中科院分区：
文献类型：
--
作者：
Hill C;Weber K
用鸡心肌残基作为抗原，可诱导出肌联蛋白特异性的鼠单克隆抗体。三种抗体T1、T3和T4在对来自鸡胸肌的多肽的免疫印迹分析中识别肌联蛋白双联体的两条带。与此相反，在鸡心肌肌原纤维的两个抗体（T1，T4）只反应的双峰，表明心脏和骨骼肌肌联蛋白之间的免疫学差异的上带。这种差异在大鼠和小鼠中更为明显。虽然所有三种抗体反应与骨骼肌肌联蛋白，T1和T4没有检测到心脏肌联蛋白，而T3反应与此肌联蛋白在免疫荧光显微镜和免疫印迹。来自各种脊椎动物的肌原纤维和冷冻组织的免疫荧光显微镜扩展了这些结果，并显示三种抗体识别不同的表位。所有三种肌联蛋白抗体装饰在新鲜制备的鸡骨骼肌和免疫电子显微镜使用天然肌球蛋白丝的肌原纤维的A-I连接处表明肌联蛋白存在于粗丝的末端。然而，在鸡心中，抗体T1和T4在I带内而不是在A-I连接处染色。这三种抗体不与任何非肌肉组织或永久性细胞系反应，也不修饰平滑肌。在鸡胚骨骼肌细胞的原代培养中，肌联蛋白首先在单核成肌细胞中以纵向条纹出现，随后在肌管的肌原纤维A-I连接处出现。
Murine monoclonal antibodies specific for titin have been elicited using a chicken heart muscle residue as antigen. The three antibodies T1, T3, and T4 recognize both bands of the titin doublet in immunoblot analysis on polypeptides from chicken breast muscle. In contrast, on chicken cardiac myofibrils two of the antibodies (T1, T4) react only with the upper band of the doublet indicating immunological differences between heart and skeletal muscle titin. This difference is even more pronounced for rat and mouse. Although all three antibodies react with skeletal muscle titin, T1 and T4 did not detect heart titin, whereas T3 reacts with this titin both in immunofluorescence microscopy and in immunoblots. Immunofluorescence microscopy of myofibrils and frozen tissues from a variety of vertebrates extends these results and shows that the three antibodies recognize different epitopes. All three titin antibodies decorate at the A-I junction of the myofibrils freshly prepared from chicken skeletal muscle and immunoelectron microscopy using native myosin filaments demonstrates that titin is present at the ends of the thick filaments. In chicken heart, however, antibodies T1 and T4 stain within the I-band rather than at the A-I junction. The three antibodies did not react with any of the nonmuscle tissues or permanent cell lines tested and do not decorate smooth muscle. In primary cultures of embryonic chicken skeletal muscle cells titin first appears as longitudinal striations in mononucleated myoblasts and later at the myofibrillar A-I junction of the myotubes.