Shibata N, Tatsumi N, Tanaka K, Okamura Y, Senda N
Biochim Biophys Acta. 1975 Aug 19;400(2):222-43. doi: 10.1016/0005-2795(75)90177-4.
The intracellular location of the binding site of antibody against purified myosin prepared from equine leucocytes was investigated in neutrophils and lymphocytes by electron microscopy using peroxidase-labelled antibody method. The myosin extracted from equine leucocytes could bind skeletal muscle F-actin and the formed complex showed the biophysical and biochemical properties and electron microscopic appearance of actomyosin. On immunodiffusion, the leucocyte myosin formed a single precipitin line with its antibody prepared in rabbits. The antibody also formed single precipitin lines with myosins from lymphocytes and thrombocytes, fusing with each other. The antibody against the leucocyte myosin did not react with myosins from skeletal or arterial smooth muscle. The specificity of the antibody was further established by determination of K+-EDTA-activated ATPase activity remained in the supernate of antigen-antibody mixture. Under electron microscope, the intracellular immunoreactive products of peroxidase labelled antibody were found in cytoplasm of neutrophils and lymphocytes incubated with antibody against leucocyte myosin, but not in neutrophils or lymphocytes treated with IgG from normal rabbits.
利用过氧化物酶标记抗体法，通过电子显微镜研究了抗马白细胞纯化肌球蛋白抗体结合位点在中性粒细胞和淋巴细胞中的细胞内定位。从马白细胞中提取的肌球蛋白能与骨骼肌F-肌动蛋白结合，形成的复合物表现出肌动球蛋白的生物物理、生化特性及电子显微镜外观。在免疫扩散实验中，白细胞肌球蛋白与其兔抗体制备的抗体形成单一沉淀线。该抗体还与淋巴细胞和血小板的肌球蛋白形成单一沉淀线，且相互融合。抗白细胞肌球蛋白抗体与骨骼肌或动脉平滑肌的肌球蛋白无反应。通过测定抗原-抗体混合物上清液中残留的K+-EDTA激活的ATP酶活性，进一步确定了抗体的特异性。在电子显微镜下，在用抗白细胞肌球蛋白抗体孵育的中性粒细胞和淋巴细胞的细胞质中发现了过氧化物酶标记抗体的细胞内免疫反应产物，而在用正常兔IgG处理的中性粒细胞或淋巴细胞中未发现。
