We show experimentally and theoretically that significant currents can be obtained with a biological ion channel, the OmpF porin of Escherichia coli, using zero-average potentials as driving forces. The channel rectifying properties can be used to pump potassium ions against an external concentration gradient under asymmetric pH conditions. The results are discussed in terms of the ionic selectivity and rectification ratio of the channel. The physical concepts involved may be applied to separation processes with synthetic nanopores and to bioelectrical phenomena. Ratchet systems have the ability of rectifying unbiased fluctuations into directed transport. 1-3 Both synthetic and biological nanochannels may show non-zero electrical currents driven by zero-average time dependent forces. [4][5][6][7] The electrical rectification is on the basis of the observed phenomena 4,5,8,9 and the potential applications concern separation processes and energy conversion. 10-12 Electrical rectification and ratchet phenomena can also be of relevance for signal averaging of weak electric fields and cell plasticity because ion channels, together with ion pumps and gap junction complexes, play a crucial role in cell communication and control. [13][14][15][16] In this Letter, we present measurements and model calculations of the rectifying properties of a biological ion channel (the outer membrane protein F, OmpF, a porin expressed in Escherichia coli bacteria 17 ). We consider the pH-controlled net current induced by zero-average oscillating potentials and the electrical pumping of potassium ions against an external concentration gradient, a phenomenon previously reported in synthetic nanopores. 4,5 We show that the complex interplay between the current rectification and the channel ionic selectivity is crucial for understanding the uphill transport of ions. The results are of relevance for the mechanisms regulating the transport through ion channels, [17][18][19] for electric signal transduction in cell cycle and embryogenesis, 14,16 and for building ionic circuits in the emerging field of nanofluidics. 6,[20][21][22][23][24][25] The set-up used in the experiments is shown schematically in Fig. 1. A single OmpF ion channel is reconstituted on a planar lipid bilayer and communicates the two halves of a conductivity cell filled with KCl solutions of concentrations c cis (side of protein addition) and c trans . pH cis and pH trans denote the corresponding pH values. A detailed description of the reconstitution procedure can be found elsewhere. 17,26 The electric potential V is positive when it is higher at the trans side of the membrane cell. An Axopatch 200B amplifier (Molecular Devices, Sunnyvale, CA) in the voltage-clamp mode is used for measuring both V and the electric current (I) through the channel. I is positive when it flows from solution trans to solution cis in Fig. 1.The OmpF porin contains both basic (positive) and acidic (negative) residues, 27 and thus, the net charge of the channel depends on the ionization state of th...