A presequence- and voltage-sensitive channel of the mitochondrial preprotein translocase formed by Tim23.

Proteins imported into the mitochondrial matrix are synthesized in the cytosol with an N-terminal presequence and are translocated through hetero-oligomeric translocase complexes of the outer and inner mitochondrial membranes. The channel across the inner membrane is formed by the presequence translocase, which consists of roughly six distinct subunits; however, it is not known which subunits actually form the channel. Here we report that purified Tim23 forms a hydrophilic, approximately 13-24 A wide channel characteristic of the mitochondrial presequence translocase. The Tim23 channel is cation selective and activated by a membrane potential and presequences. The channel is formed by the C-terminal domain of Tim23 alone, whereas the N-terminal domain is required for selectivity and a high-affinity presequence interaction. Thus, Tim23 forms a voltage-sensitive high-conductance channel with specificity for mitochondrial presequences.

A Ca(2+)- and voltage-modulated flagellar ion channel is a component of the mechanoshock response in the unicellular green alga Spermatozopsis similis.

In flagellate green algae, behavioral responses to photo- and mechanoshock are induced by different external stimuli within 10-15 ms. In the accompanying changes in flagella beat, Ca(2+) has important regulatory roles. Although the axonemal Ca(2+) responsive elements are well characterized, analyses of flagellar channels involved in Ca(2+) signalling as well as other ion channels at the single-channel level were not yet conducted in green algae. To gain a further understanding of these important signaling elements in movement responses, intact flagella of Spermatozopsis similis were isolated and characterized and the solubilized flagellar membrane proteins were reconstituted into liposomes. We observed three types of channel activity, two of which were weakly anion and cation-selective and in the high-conductance regime typical for porin-like solute channels. The dominating channel activity was a voltage dependent, rectifying, low conductance (Lambda=80 pS in 50 mM KCl) cation-selective channel modulated by, and highly permeable to, Ca(2+) ions (SFC1: Spermatozopsis flagellar cation channel 1). Depolarizations necessary to activate SFC1 probably only occur in vivo during avoidance reactions of this alga. Ca(2+)-activation of SFC1 points to a direct link to Ca(2+)-mediated signaling pathway(s) in the flagella. Both the response to mechanoshock and SFC1 activity were inhibited by Gd(3+) and Ba(2+), thus supporting our assumption that SFC1 represents a major flagellar ion channel involved in this green algal avoidance reaction.