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J. Biol. Chem., Vol. 277, Issue 23, 20270-20276, June 7, 2002
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From the Departments of Medicine and Biochemistry, McGill
University, Montreal, Quebec H3G 1Y6, Canada
The two variants of the
Distinct Regulatory Effects of the Na,K-ATPase
Subunit*
subunit of the rat
renal sodium pump, a and b, have
similar effects on the Na,K-ATPase. Both increase the affinity
for ATP due to a shift in the enzyme's E1 
E2 conformational equilibrium toward E1. In
addition, both increase K+ antagonism of cytoplasmic
Na+ activation. To gain insight into the structural basis
for these distinct effects, extramembranous N-terminal and C-terminal
mutants of were expressed in rat 
1-transfected HeLa cells. At
the N terminus, the variant-distinct region was deleted (N
7) or
replaced by alanine residues (N7A). At the C terminus, four
(aC4) or ten (aC10) residues were
deleted. None of these mutations abrogates the
K+/Na+ antagonism as evidenced in a similar
increase in K'Na seen at high (100 mM) K+ concentration. In contrast, the
C-terminal as well as N-terminal deletions (N7,
aC4, and aC10) abolished the
decrease in K'ATP seen with wild-type
a or b. It is concluded that different regions of the chain mediate the distinct functional effects of
, and the effects can be long-range. In the transmembrane region,
the impact of G41R replacement was analyzed since this mutation is
associated with autosomal dominant renal Mg2+-wasting in
man (Meij, I. C., Koenderink, J. B., van Bokhoven, H.,
Assink, K. F. H., Groenestege, W. T., de Pont, J. J. H. H. M., Bindels, R. J. M., Monnens, L. A. H., Van
den Heuvel, L. P. W. J., and Knoers, N. V. A. M. (2000) Nat.
Genet. 26, 265-266). The results show that Gly-41 
Arg
prevents trafficking of but not 
pumps to the cell surface
and abrogates functional effects of on pumps. These
findings underscore a potentially important role of in affecting
solute transport, in this instance Mg2+ reabsorption,
consequent to its primary effect on the sodium pump.
*
This work was supported by operating grants from the
Canadian Institutes of Health Research (MT-3876) and the Kidney
Foundation of Canada.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Montreal General
Hospital, 1650 Cedar Ave., Montreal, Quebec H3G 1A4, Canada. Tel.:
514-934-1934 (ext. 44501); Fax: 514-934-8332; E-mail: Rhoda. Blostein{at}mcgill.ca.
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