Cellular assays for measuring dynamin activity in muscle cells

Abstract

Dynamin is one of the best-studied membrane fission machineries, which mediates endocytic vesicle pinch-off from the plasma membrane. Among the three dynamin isoforms encoded in mammalian genome, dynamin-2 is the ubiquitously expressed isoform and leads to human muscular or neuronal diseases when mutants causing hyperactivity or hypoactivity of its membrane fission activity occur. While transferrin uptake is the most commonly used assay to measure dynamin activity in cultured cells, here we provide two different methods to quantitatively examine the activity of dynamin in myoblasts and myotubes, i.e., Bin1-tubule vesiculation and glucose transporter 4 fractionation assays, respectively. These methods could provide a quantitative measurement of dynamin activity in both differentiated and undifferentiated myoblasts.