CONNECTIN, AN ELASTIC PROTEIN OF MUSCLE - IDENTIFICATION OF TITIN WITH CONNECTIN
CONNECTIN, AN ELASTIC PROTEIN OF MUSCLE - IDENTIFICATION OF TITIN WITH CONNECTIN
复制标题
DOI：
10.1093/oxfordjournals.jbchem.a133249
复制
发表时间：
1981-01-01
影响因子：
2.7
通讯作者：
KUWANO, Y
中科院分区：
文献类型：
--
作者：
MARUYAMA, K;KIMURA, S;KUWANO, Y
当兔和鸡骨骼肌的整个肌纤维或肌原纤维直接溶解在热SDS [十二烷基硫酸钠]溶液中时，可以通过凝胶过滤分离出一种称为肌联蛋白的非常高分子量的蛋白质。连接蛋白是肌肉的弹性蛋白，可以通过类似的方法从完全提取的肌肉残留物中分离。用2-3%聚丙烯酰胺凝胶电泳、间接免疫荧光技术测定肌原纤维的氨基酸组成和定位，结果表明肌联蛋白和连接蛋白是相同的。在室温下储存几天后，连接蛋白在SDS溶液中不稳定; SDS凝胶电泳上连接蛋白的双重带变得弥散并最终消失。连接蛋白集中在肌原纤维的A-I连接区周围，尽管它存在于除Z线之外的整个肌节中。去除肌球蛋白后，用连接蛋白荧光抗体处理时，A-I连接处仍有荧光。在KI提取的肌原纤维中，聚集在Z线两侧的材料被强烈染色，相邻Z线之间存在荧光丝，但Z线完全不染色。
When whole muscle fibers or myofibrils of rabbit and chicken skeletal muscles are directly solubilized in hot SDS [sodium dodecylsulfate] solution, a very high MW protein called titin can be isolated by gel filtration. Connectin, an elastic protein of muscle, can be isolated by a similar method from thoroughly extracted muscle residues. Studies of electrophoretic mobility on 2-3% polyacrylamide gel electrophoresis, amino acid composition and localization in myofibrils determined by the indirect immunofluorescence technique showed that titin and connectin are identical. Connectin was unstable in SDS solution on storage for a few days at room temperature; the doublet band of connectin on SDS gel electrophoresis became diffuse and eventually disappeared. Connectin was concentrated around the A-I junction region of a myofibril, although it was present in an entire sarcomere except for the Z lines. On removal of myosin, the A-I junction was still fluorescent, when treated with fluorescent antibody against connectin. In the KI-extracted myofibril, materials accumulated on both sides of the Z lines were strongly stained and there were fluorescent filaments between the neighboring Z lines, but the Z lines were not stained at all.