Price M G
J Cell Biol. 1987 May;104(5):1325-36. doi: 10.1083/jcb.104.5.1325.
The cytoskeletons of mammalian striated and smooth muscles contain a pair of high molecular weight (HMW) polypeptides of 220,000 and 200,000 mol wt, each with isoelectric points of about 5 (Price, M. G., 1984, Am. J. Physiol., 246:H566-572) in a molar ratio of 1:1:20 with desmin. The HMW polypeptides of mammalian muscle have been named "skelemins," because they are in the insoluble cytoskeletons of striated muscle and are at the M-discs. I have used two-dimensional peptide mapping to show that the two skelemin polypeptides are closely related to each another. Polyclonal antibodies directed against skelemins were used to demonstrate that they are immunologically distinct from talin, fodrin, myosin heavy chain, synemin, microtubule-associated proteins, and numerous other proteins of similar molecular weight, and are not oligomers of other muscle proteins. Skelemins appear not to be proteolytic products of larger proteins, as shown by immunoautoradiography on 3% polyacrylamide gels. Skelemins are predominantly cytoskeletal, with little extractable from myofibrils by various salt solutions. Human, bovine, and rat cardiac, skeletal, and smooth muscles, but not chicken muscles, contain proteins cross-reacting with anti-skelemin antibodies. Skelemins are localized by immunofluorescence at the M-lines of cardiac and skeletal muscle, in 0.4-micron-wide smooth striations. Cross sections reveal that skelemins are located at the periphery of the M-discs. Skelemins are seen in threads linking isolated myofibrils at the M-discs. There is sufficient skelemin in striated muscle to wrap around the M-disc about three times, if the skelemin molecules are laid end to end, assuming a length-to-weight ratio similar to M-line protein and other elongated proteins. The results indicate that skelemins form linked rings around the periphery of the myofibrillar M-discs. These cytoskeletal rings may play a role in the maintenance of the structural integrity of striated muscle throughout cycles of contraction and relaxation.
哺乳动物横纹肌和平滑肌的细胞骨架含有一对分子量为220,000和200,000道尔顿的高分子量(HMW)多肽,其等电点均约为5(普赖斯,M.G.,1984年,《美国生理学杂志》,246:H566 - 572),与结蛋白的摩尔比为1:1:20。哺乳动物肌肉的HMW多肽被命名为“骨骼蛋白”,因为它们存在于横纹肌的不溶性细胞骨架中且位于M盘。我利用二维肽图谱分析表明这两种骨骼蛋白多肽彼此密切相关。针对骨骼蛋白的多克隆抗体被用于证明它们在免疫上与踝蛋白、血影蛋白、肌球蛋白重链、伴肌动蛋白、微管相关蛋白以及许多其他分子量相似的蛋白质不同,并且不是其他肌肉蛋白的寡聚体。如在3%聚丙烯酰胺凝胶上进行免疫放射自显影所示,骨骼蛋白似乎不是较大蛋白质的蛋白水解产物。骨骼蛋白主要存在于细胞骨架中,用各种盐溶液从肌原纤维中几乎提取不出来。人、牛和大鼠的心肌、骨骼肌和平滑肌含有与抗骨骼蛋白抗体发生交叉反应的蛋白质,但鸡肌肉中没有。通过免疫荧光法,骨骼蛋白定位于心肌和骨骼肌的M线处,呈0.4微米宽的平滑条纹。横截面显示骨骼蛋白位于M盘的周边。在连接M盘处分离的肌原纤维的细丝中可以看到骨骼蛋白。如果骨骼蛋白分子首尾相连,假设其长度与重量比类似于M线蛋白和其他伸长蛋白,横纹肌中的骨骼蛋白足以围绕M盘约三圈。结果表明,骨骼蛋白在肌原纤维M盘的周边形成连接环。这些细胞骨架环可能在横纹肌收缩和舒张的整个周期中维持其结构完整性方面发挥作用。