Birk Jesper B, Wojtaszewski Jørgen F P
Section of Molecular Physiology, Department of Nutrition, Exercise and Sports, Faculty of Science, University of Copenhagen, Copenhagen, Denmark.
Methods Mol Biol. 2018;1732:203-213. doi: 10.1007/978-1-4939-7598-3_13.
The 5'-AMP-activated protein kinase is a complicated enzyme consisting of three different subunits, each of which is expressed as two or three isoforms. This gives the possibility of 12 different heterotrimeric complexes, which could have diverse functions within the cell. To map out which of these complexes are present and to what extent in skeletal muscle, we have used the immunoprecipitation technique and analyzed both the precipitates and the remaining supernatants for coprecipitation of complex partners. We have fine-tuned this method to give the best results on lysates from the skeletal muscle, liver, and heart muscle from mouse to man.
5'-AMP激活蛋白激酶是一种由三个不同亚基组成的复杂酶，每个亚基又以两种或三种同工型表达。这就产生了12种不同异源三聚体复合物的可能性，它们在细胞内可能具有多种功能。为了确定这些复合物中哪些存在于骨骼肌中以及其存在程度，我们使用了免疫沉淀技术，并分析沉淀物和剩余上清液中复合物伙伴的共沉淀情况。我们对该方法进行了微调，以便在从小鼠到人类的骨骼肌、肝脏和心肌裂解物上获得最佳结果。
