4. Conclusions
Purified, water-soluble ICln protein forms ion channels by spontaneously incorporating into artificial lipid bilayers. Site-directed mutation experiments revealed E41 and D49 as the amino acids responsible for the Ca2+-dependence of the channels’ ion selectivity and G49 to be part of the putative nucleotide binding site of the protein. H64 could be pinpointed within the ion conducting pathway of ICln. In the present study, cysteine mutagenesis and Cd2+ block was used to further investigate the pore structure of the membrane spanning β-barrel formed by ICln, as proposed by the computer model and NMR studies. The results confirm that the amino acids G49 and S55 have access to the pore lumen of reconstituted ICln channels, as predicted by the model, and that S50 and L52 are facing the hydrophobic membraneous phase. According to the lack of Cd2+-block of ICln-G51C channels, the proposed ICln channel model has to be revised insofar as G51 seems not to face the pore lumen or is not accessible to Cd2+.
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Jakab, M. et al. (2004). Probing of the ICln Channel Pore by Cysteine Mutagenesis and Cadmium-Block. In: Lauf, P.K., Adragna, N.C. (eds) Cell Volume and Signaling. Advances in Experimental Medicine and Biology, vol 559. Springer, Boston, MA . https://doi.org/10.1007/0-387-23752-6_9
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