Dysferlin and its role in cardiac disease

To explore the proteomic interactome of Dysferlin in the heart, we used recombinant expression of N-terminally Twin-Strep II-tagged full-length human Dysferlin to establish an in vitro binding assay. Isolated mouse heart membrane fractions were incubated with Dysferlin-coupled beads under four different detergent conditions. Dysferlin pull-down eluates were analyzed by label-free quantification using an ion mobility-enhanced data-independent acquisition (DIA) workflow with alternating low and elevated energy (referred to as UDMSE).

为探究心脏中Dysferlin的蛋白质组学互作组，我们通过重组表达N端带有Twin-Strep II标签的全长人源Dysferlin，建立了体外结合实验体系。将分离获得的小鼠心脏膜组分与偶联有Dysferlin的磁珠在四种不同去污剂条件下共孵育。随后采用结合高低能量交替模式的离子迁移增强型数据非依赖采集（DIA）分析流程（亦称UDMSE）结合无标记定量技术，对Dysferlin下拉实验的洗脱液进行分析。