Lehman W, Denault D, Marston S
Department of Physiology, Boston University School of Medicine, MA.
Biochim Biophys Acta. 1993 Nov 10;1203(1):53-9. doi: 10.1016/0167-4838(93)90035-p.
Caldesmon and tropomyosin can be selectively and quantitatively extracted from vascular and visceral smooth muscle following heat treatment; all other smooth muscle proteins are precipitated by this procedure. Estimates of the caldesmon/tropomyosin molar ratio in heat-extracts determined by SDS-PAGE densitometry are 1 caldesmon:5.1-5.3 tropomyosin for rabbit and sheep aorta, and 1 caldesmon:5.9 tropomyosin for rabbit stomach and chicken gizzard. If the assumption is made that tropomyosin serves as a true reference of thin-filament content in intact muscle, it follows that the relative caldesmon contents in the above tissues are similar to each other. Caldesmon in heat extracts was identified by Western blotting, by its anomalous migration on several different SDS-PAGE systems and by its position on two-dimensional PAGE. Values of caldesmon contents in unfractionated total tissue homogenates were found to be similar to those cited above. Smooth muscles contain different thin-filament classes and only one type appears to possess caldesmon. By comparing values for the molar composition of caldesmon-specific filaments (1 caldesmon:2 tropomyosin:14 actin) with the values above determined for intact tissue, we conclude that the caldesmon filaments account for approx. 35-45% of the total thin-filament pool in arterial smooth muscle and slightly less in visceral muscles.
热处理后,钙调蛋白和原肌球蛋白可从血管和内脏平滑肌中被选择性地定量提取;此过程会使所有其他平滑肌蛋白沉淀。通过SDS-PAGE光密度法测定的热提取物中钙调蛋白/原肌球蛋白的摩尔比,兔和羊主动脉为1份钙调蛋白:5.1 - 5.3份原肌球蛋白,兔胃和鸡胗为1份钙调蛋白:5.9份原肌球蛋白。如果假设原肌球蛋白可作为完整肌肉中细肌丝含量的真实参照,那么上述组织中钙调蛋白的相对含量彼此相似。热提取物中的钙调蛋白通过蛋白质免疫印迹法、在几种不同SDS-PAGE系统上的异常迁移以及在双向PAGE上的位置得以鉴定。未分级的全组织匀浆中钙调蛋白的含量与上述值相似。平滑肌含有不同类型的细肌丝,似乎只有一种类型含有钙调蛋白。通过将钙调蛋白特异性细丝的摩尔组成值(1份钙调蛋白:2份原肌球蛋白:14份肌动蛋白)与上述完整组织测定值进行比较,我们得出结论,钙调蛋白细丝约占动脉平滑肌中细肌丝总量的35 - 45%,在内脏肌肉中所占比例略低。