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J Biol Chem, Vol. 274, Issue 16, 11376-11382, April 16, 1999
From the Department of Biological Sciences, By using differential mRNA display to monitor
the molecular alterations associated with adaptation of euryhaline eels
to different salinities, we identified a cDNA fragment strongly
induced in seawater eel gills. Cloning of a full-length cDNA and
its expression in COS-7 cells indicated that the clone codes for an
inward rectifier K+ channel (eKir) of 372 amino acid
residues, which has two transmembrane segments and a typical
pore-forming region (H5). Only low sequence similarities are present,
except the H5 region, compared with other members of the inward
rectifier K+ channel family (Kir). Consistent with this
divergence in the amino acid sequence, a phylogenetic analysis
indicated early divergence and independent evolution of eKir from other
members; it is only distantly related to the Kir5.0 subfamily members.
RNase protection analysis showed that eKir is highly expressed in the
seawater eel gill, kidney, and posterior intestine but very weakly in
freshwater eels. Immunohistochemistry of gill sections revealed dense
localization of eKir in the chloride cells. Immunoelectron microscopy
indicated that eKir is mainly present in the microtubular system in the chloride cell. This location and its salt-inducible nature suggest that
the eKir channel cloned here is a novel member of the Kir5.0 subfamily
of the Kir family and is implicated in osmoregulation.
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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