Adult skeletal muscles express two Na,K-ATPase isoforms, α1 and α2. α2, the sole isoform expressed in the transverse tubules, comprises up to 90% of total Na,K-ATPase. α2 in resting muscle operates well below its maximum turnover rate; but is rapidly stimulated, by yet unknown mechanisms, during muscle contraction, helping maintaining excitation and resist fatigue. We tested the hypothesis that α2 activity may be regulated by extracellular K over the physiological range of K concentrations that occur in the t-tubules. Na,K-ATPase driven current (Ipump) was measured in enzymatically isolated FDB fibers voltage clamped at −90 mV using a two microlectrode amplifier. Ipump was identified as a ouabain- and temperature-sensitive outward current activated by Ko in fibers rendered electrically passive. Intracellular solutions (filling both electrodes) contained (mM): 1EGTA, 0.5CaCl2, 5MgCl2, 20MOPS, 5gluthathion-H2, 90K-asparte, and 5ATP-Na2, 5-phosphocreatine-Na2, and 30Na-aspartate to promote forward pump cycling. External solutions contained (mM): 2CaCl2, 1MgCl2, 10glucose, 10MOPS; and inversely varying amounts of KCl (0-40) and NaCL (154-114) to maintain osmolarity. TTX (400nM), nifedipine (20 μM), Ba (1mM) and 9-anthracene-carboxilic-acid (200μM) were added to block main ionic currents. All solutions had pH=7.4 and 300mOmal/kgH2O. Ipump was activated in a concentration-dependent manner by raising [K]o from 0 to 40mM, and eliminated by washing out K. Ipump was prevented by pretreatment with 10μM ouabain or inhibited by addition of ouabain, and was steeply temperature-dependent. Maximal Ipump vs. [K]o was fitted to obtain the Km for pump activation by K. Results demonstrate that α2 in skeletal muscle is regulated by changes in extracellular K over tens of mM, suggesting that this isoform is adapted to respond to expected demands of Na/K transport in the t-tubules during sustained activity. (Supported by the National institutes of Health, USA)