Mogami K, Fujita S C, Hotta Y
J Biochem. 1982 Feb;91(2):643-50. doi: 10.1093/oxfordjournals.jbchem.a133736.
When proteins of whole Drosophila thorax were analyzed by two-dimensional gel electrophoresis, 186 spots were detected by protein staining with Coomassie brilliant blue R-250. Two methods were developed to identify proteins which exist in indirect flight muscle (IFM) and its myofibrils. 1) A whole fly was freeze-dried in a dry ice-acetone mixture, and indirect flight muscle fibers were cleanly dissected out from the thorax. The muscle cells and the rest of the thorax were analyzed separately. The muscle contained 146 polypeptides, of which 12 were not detected elsewhere. 2) Flies were frozen in liquid nitrogen and shaken vigorously so that their thoraces broke off from heads and abdomens. The thoraces were separated from the rest by sieving and centrifugation. After homogenization of the thorax, myofibrils were prepared by centrifugation in a discontinuous sucrose density gradient. The myofibril fraction contained at least 20 proteins. There were two types of actin (II and III), myosin heavy chain, tropomyosin and paramyosin. Nine of the other myofibrillar proteins were specific to this muscle.
当用二维凝胶电泳分析果蝇整个胸部的蛋白质时，用考马斯亮蓝R - 250进行蛋白质染色检测到了186个斑点。开发了两种方法来鉴定存在于间接飞行肌（IFM）及其肌原纤维中的蛋白质。1）将整只果蝇在干冰 - 丙酮混合物中冷冻干燥，然后从胸部干净地解剖出间接飞行肌纤维。分别对肌肉细胞和胸部的其余部分进行分析。该肌肉含有146种多肽，其中12种在其他地方未被检测到。2）将果蝇在液氮中冷冻并剧烈摇晃，使其胸部与头部和腹部分离。通过筛分和离心将胸部与其余部分分开。胸部匀浆后，通过在不连续蔗糖密度梯度中离心制备肌原纤维。肌原纤维部分至少含有20种蛋白质。有两种肌动蛋白（II型和III型）、肌球蛋白重链、原肌球蛋白和副肌球蛋白。其他9种肌原纤维蛋白是这种肌肉特有的。
