Phillips C L, Yamakawa K, Adelstein R S
Laboratory of Molecular Cardiology National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA.
J Muscle Res Cell Motil. 1995 Aug;16(4):379-89. doi: 10.1007/BF00114503.
Previously, we reported the sequence of cDNA clones encoding amino acids 63 through 723 of the human nonmuscle myosin heavy chain-B isoform. In this paper, we present the derived sequence of the remaining 1303 amino acids along with 5' and 3' untranslated sequences. We made use of the differences between the derived nonmuscle myosin heavy chain-A and -B amino acid sequences to raise isoform-specific antibodies. Immunoblot analysis reveals a differential expression of both myosin heavy chain isoforms in a variety of human adult and foetal tissues and cells. When extracts of human adult aorta were subjected to gel electrophoresis, two distinct Coomassie Blue-stained bands and a fused band were seen migrating at approximately 200 kDa. These bands can be detected with four different specific antibodies recognizing the two different smooth muscle myosin heavy chain isoforms (204 kDa and 200 kDa) and the two different nonmuscle myosin heavy chain isoforms (A and B). Using immunohistochemistry, we confirmed the presence of the four different isoforms in adult and foetal aortas.
此前，我们报道了编码人非肌肉肌球蛋白重链 -B 亚型第 63 至 723 个氨基酸的 cDNA 克隆序列。在本文中，我们展示了其余 1303 个氨基酸的推导序列以及 5' 和 3' 非翻译序列。我们利用推导的非肌肉肌球蛋白重链 -A 和 -B 氨基酸序列之间的差异来制备亚型特异性抗体。免疫印迹分析揭示了两种肌球蛋白重链亚型在多种人类成人和胎儿组织及细胞中的差异表达。当对人类成人主动脉提取物进行凝胶电泳时，可见两条明显的考马斯亮蓝染色带和一条融合带在约 200 kDa 处迁移。这些条带可用四种不同的特异性抗体检测到，这些抗体分别识别两种不同的平滑肌肌球蛋白重链亚型（204 kDa 和 200 kDa）以及两种不同的非肌肉肌球蛋白重链亚型（A 和 B）。通过免疫组织化学，我们证实了在成人和胎儿主动脉中存在这四种不同的亚型。
