Le Peuch C J, Demaille J G, Pechere J F
Biochim Biophys Acta. 1978 Nov 20;537(1):153-9. doi: 10.1016/0005-2795(78)90610-4.
A two-dimensional radioelectrophoretic method is described, by which parvalbumins from minute biopsy samples (approx. 50 mg) can be detected and quantitated by their 45Ca2+-binding properties. In the first dimension, parvalbumins are purified by sieving through a gradient polyacrylamide gel and collected at the bottom of the electrophoresis tubes. The second dimension is a disc electrophoresis in the presence of 45Ca2+. Parvalbumins can thus be identified and quantitated by three criteria: low molecular weight, acidic character and calcium-binding properties, since they are never exposed to denaturing conditions. Validity of the technique was demonstrated on carp myogen, and on extracts from rabbit psoas and heart muscles. Application of this method to the shrew fast beating myocardium shows that it does contain parvalbumin, in agreement with the proposed role of soluble relaxing factor (Pechère et al. (1977) FEBS Lett. 75, 111--1141. When applied to human muscle biopsies, radioelectrophoresis points to an uneven distribution of parvalbumin among different skeletal muscles. For the human limb muscles tested in this study, the parvalbumin content is similar to that of rabbit psoas muscle.
本文描述了一种二维放射电泳方法，通过该方法可利用小活检样本（约50毫克）中的小清蛋白与45Ca2+的结合特性对其进行检测和定量。在第一维中，小清蛋白通过梯度聚丙烯酰胺凝胶筛分进行纯化，并收集在电泳管底部。第二维是在45Ca2+存在下的圆盘电泳。由于小清蛋白从未暴露于变性条件下，因此可通过三个标准对其进行鉴定和定量：低分子量、酸性特征和钙结合特性。该技术在鲤鱼肌原蛋白以及兔腰大肌和心肌提取物上得到了验证。将该方法应用于鼩鼱快速跳动的心肌表明，其确实含有小清蛋白，这与可溶性舒张因子的推测作用一致（佩谢尔等人（1977年），《欧洲生物化学学会联合会快报》75，111 - 114）。当应用于人体肌肉活检时，放射电泳表明小清蛋白在不同骨骼肌中的分布不均匀。对于本研究中测试的人体肢体肌肉，小清蛋白含量与兔腰大肌相似。
