Letters to the Editor

Department of Pharmacology, University of Cambridge, Cambridge CB2 1QJ, UK The following is the abstract of the article discussed in the subsequent letter: Blazer-Yost, Bonnie L., and Sandy I. Helman. The amiloride-sensitive epithelial Na + channel: binding sites and channel densities. Am. J. Physiol. 272 ( Cell Physiol. 41): C761-C769, 1997. The amiloride-sensitive Na + channel found in many transporting epithelia plays a key role in regulating salt and water homeostasis. Both biochemical and biophysical approaches have been used to identify, characterize, and quantitate this important channel. Among biophysical methods, there is agreement as to the single-channel conductance and gating kinetics of the highly selective Na + channel found in native epithelia. Amiloride and its analogs inhibit transport through the channel by binding to high-affinity ligand-binding sites. This characteristic of high-affinity binding has been used biochemically to quantitate channel densities and to isolate presumptive channel proteins. Although the goals of biophysical and biochemical experiments are the same in elucidating mechanisms underlying regulation of Na + transport, our review highlights a major quantitative discrepancy between methods in estimation of channel densities involved in transport. Because the density of binding sites measured biochemically is three to four orders of magnitude in excess of channel densities measured biophysically, it is unlikely that high-affinity ligand binding can be used physiologically to quantitate channel densities and characterize the channel proteins.