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J Biol Chem, Vol. 274, Issue 47, 33183-33185, November 19, 1999
From the Laboratory of Membrane Biology, Neuroscience Center,
Massachusetts General Hospital, Charlestown, Massachusetts 02129
The Na+,K+-ATPase
catalyzes the active transport of ions. It has two necessary subunits,
The renal control of Na+ and K+ balance is
complex and entails ensembles of apical and basolateral transporters
that play specialized roles in different segments of the nephron. One
of the most physiologically important transporters is the Na,K-ATPase,
or sodium pump, which is crucial for the absorptive, secretory, and
concentrating capacity of the kidney. Small changes in Na,K-ATPase ion
affinities can have important physiological effects both directly on
transepithelial ion and fluid transport and indirectly through the ion
gradients used by other transporters (1, 2). In permeabilized
microdissected nephron segments, Na,K-ATPase has increased affinity for
Na+ in thick ascending limb compared with proximal
convoluted tubule and has still higher affinity in cortical collecting
tubule (3-6). Although isoforms of A small single-span membrane protein, member of a protein family that
has ion channel activity in oocytes, copurifies with renal Na,K-ATPase
Enzyme Preparations--
Na,K-ATPase was purified from rat, pig,
and dog kidney by SDS extraction (21), which produces membrane bound,
active enzyme. Crude membranes from renal cell lines and transfectants
were treated with SDS at 0.56 mg/mg protein and sedimented on 7-30%
sucrose gradients. Specific activities of the Na,K-ATPase from rat
kidney and NRK-52E cells were 1200-1500 and 120-150 µmol of
Pi/mg/hr, respectively.
Gels--
Electrophoresis was in
SDS-Tricine1 gels (22).
Proteins were transferred to nitrocellulose, incubated with antibodies,
and detection was with chemiluminescence (23).
Antibody to ATP Hydrolysis--
Na,K-ATPase activity was measured in media
containing 3 mM Tris-ATP, 3 mM
MgCl2, 30 mM histidine, pH 7.3, and various
concentrations of Na+ (0-20 mM) with
[K+] fixed at 20 mM, or various
concentrations of K+ (0-4 mM) with
[Na+] fixed at 140 mM. Reactions were
performed for 30 min at 37 °C with and without 3 mM
ouabain, and ouabain-sensitive Pi release was
measured colorimetrically.
Transfections--
cDNA for the Immunofluorescence--
Cryostat sections were made from rat
kidney fixed with periodate-lysine-paraformaldehyde. Sections were
double-labeled with mouse anti- Properties of
To determine whether Post-translational Modification of
Hydroxylamine treatment was used to determine whether the modification
was because of acylation of the protein (23, 26). At basic pH,
hydroxylamine shifted the slower-migrating band but left the
faster-migrating band unchanged (Fig. 3b). At neutral pH
neither band was affected (not shown), and there is no Asn-Gly in the
sequence for hydroxylamine to cleave (27). This suggests a hydroxyester
linkage, possibly fatty acid acylation, to Ser or Thr. There are three
Ser and Thr residues in the N-terminal segment that are conserved in
Localization of The most notable consequence of NRK-52E is morphologically heterogeneous, lending credibility to the
idea that subclones may have different phenotypes with respect to The anatomical distribution of Pathways that alter ion affinities of kidney Na,K-ATPase consequent to
hormone treatment or electrolyte dietary restriction have been reported
(4, 5, 28, 29). Their molecular mechanisms are still obscure, however,
and could entail the expression and/or modification of We would like to thank Robert Mercer, Steven
Karlish, and Kaethi Geering for samples of anti- *
This work was supported by National Institutes of Health
Grants NS27653 and HL36271.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.
The abbreviations used are:
Tricine, N-[2-hydroxy-1,1-bis(hydroxymethyl)ethyl]glycine;
RT-PCR, reverse transcriptase polymerase chain reaction.
COMMUNICATION
The 
Subunit Modulates Na+ and K+
Affinity of the Renal Na,K-ATPase*
ABSTRACT
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
and 
, but in kidney it is also associated with a 7.4-kDa
protein, the 
subunit. Stable transfection was used to determine the
effect of on Na,K-ATPase properties. When isolated from either
kidney or transfected cells, had lower affinities for both
Na+ and K+ than . A post-translational
modification of selectively eliminated the effect on
Na+ affinity, suggesting three configurations (,
, and *) conferring different stable properties to
Na,K-ATPase. In the nephron, segment-specific differences in
Na+ affinity have been reported that cannot be explained by
the known and subunit isoforms of Na,K-ATPase.
Immunofluorescence was used to detect in rat renal cortex. Cortical
ascending limb and some cortical collecting tubules lacked ,
correlating with higher Na+ affinities in those segments
reported in the literature. Selective expression in different segments
of the nephron is consistent with a modulatory role for the subunit
in renal physiology.
INTRODUCTION
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
and can have different ion
affinities (7), isoforms unique to the nephron segments with higher
affinity have not been detected (8-12), leaving the mechanism unexplained.
and subunits and appears as a doublet on gels (13, 14). Known
as the subunit, it is not present in all tissues, and its role in
the Na,K-ATPase has long been an enigma (15-17). As an ancillary
subunit it is likely to play a modulatory role. It has been reported to
increase ATP affinity when expressed in mammalian cells (18, 19) and to
affect the voltage sensitivity of K+ activation when
expressed in oocytes (20). Here evidence is presented for its role in
the control of the most basic property of Na,K-ATPase, the affinity for
Na+ and K+. Most significantly, analysis of
independent clones points to a functional role for post-translational
modification of the subunit.
EXPERIMENTAL PROCEDURES
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
--
A rabbit antiserum (RCT-G1) was raised
against the peptide corresponding to the last 14 amino acids
(CGGSKKHRQVNEDEL) of rat , conjugated to keyhole limpet
hemocyanin (KLH).
subunit was obtained by
RT-PCR from total rat kidney RNA (CLONTECH, Palo
Alto, CA). The following primers were based on a nucleotide sequence
for rat in the GenBankTM dbEST data base (AA801241)
plus EcoRI and BamHI restriction sites for
unidirectional cloning: forward primer, 5'-GGAATTCGTGGCTGGGGAAATGAC-3'; reverse primer, 5'-CGCGGATCCCAGCTCATCTTCATTGAC-3'.
Gel-purified DNA was ligated into pIRES vector
(CLONTECH). Several clones containing the
full-length cDNA of were verified by PCR and nucleotide sequencing. The N-terminal protein sequence is predicted to be MTELSANHGGS, corresponding to the corrected sequence reported by Minor
et al. (14) and different from that deposited in
GenBankTM (X70062). Transfection was with cationic
liposomes (Clonfectin, CLONTECH). Antibiotic (G418)
was added after 48 h, and resistant colonies formed in 15-20 days.
1 (McK1, 1:4) and rabbit anti-
(RCT-G1, 1:500), stained with Cy3-conjugated anti-mouse and fluorescein
isothiocyanate-conjugated anti-rabbit secondary antibodies, and
examined with a Bio-Rad MRC 1024 Laser Sharp confocal microscope.
RESULTS
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
Transfectants--
While Na,K-ATPase from kidney
contains , this is not true of mammalian kidney cell lines (18).
NRK-52E, an epithelial line of rat kidney cells, expressed the same
Na,K-ATPase 1 and 1 isoforms found in rat kidney, but could
not be detected with specific antibodies (Fig.
1) or by RT-PCR (not shown). The
Na+ and K+ affinities of partially purified
Na,K-ATPase with and without were compared (Table
I, top). The apparent affinities for both ions were substantially higher (1.5-2-fold) in enzyme from NRK-52E (11) than from renal medulla (11). Two other renal cell
lines, LLC-PK1 (pig) and MDCK (dog), also lacked (not shown), and
again cell-derived enzyme without had higher affinities for
Na+ than renal enzyme with it (Table I, top).
View larger version (47K):
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Fig. 1.
Detection of .
Western blots stained for 1 and of Na,K-ATPase partially
purified from rat renal medulla (lane 1), NRK-52E control
cells (lane 2), NRK-52E transfected with empty vector
(lane 3), and two representative clones transfected with (lanes 4 and 5). The blot was cut so that the high molecular
weight region could be stained with antibody McK1 (1) and the
peptide region with antiserum RCT-G1 ().
Ion affinities for ATP hydrolysis by Na,K-ATPase with and without
was responsible for the kinetic difference,
stable transfectants of NRK-52E were generated. Expression of was
detected on Western blots (Fig. 1). By densitometric analysis, the
level was 15-20% of the level in the kidney enzyme, normalized to the
amount of 1. Two groups of clones were distinguished by the mobility
of : those expressing a doublet (as in kidney preparations) and
those expressing only the band with slower mobility. The distinction
between clones proved to be critical for the kinetic properties of the
Na,K-ATPase. The clones that expressed a doublet showed reduced
affinity for Na+ as well as K+, while the
clones with only one band showed reduced affinity for K+
only (Fig. 2). This relationship held
true in three independent clones with doublets and three clones with
single bands. The ion affinities for all of the tested clones are
compared in Table I, bottom.
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Fig. 2.
Na+ and
K+ affinities were altered by
and its posttranslational modification.
Hydrolysis of ATP was measured as a function of either Na+
or K+. Partially purified enzyme from mock-transfected
NRK-52E (
) or clones expressing either the doublet (
) or the
single band (
) were tested. Data from Na,K-ATPase purified from rat
renal medulla (
, dashed lines) are shown on each graph
for comparison. a, Na+ affinity; b,
K+ affinity. Expression of either form of altered
K+ affinity to resemble that of kidney-derived enzyme,
while only the clones expressing the doublet had altered
Na+ affinity.
--
In the literature,
doublets of mammalian appeared during in vitro synthesis
in the presence of pancreatic microsomes (17), but not in their absence
(20), which suggests that there might be post-translational processing
in the endoplasmic reticulum. The subunit does not have a signal
sequence, but trypsin treatment of pig kidney right-side-out sealed
microsomal vesicles reduced the doublet to a single band, consistent
with a cleavage site near the extracellular N terminus (18). We
digested right-side-out rat renal medulla vesicles with trypsin (24).
Both bands of the doublet were reduced to one smaller band, from
7.6 and 6.8 kDa to 5.6 kDa (Fig.
3a), predicting cleavage at
Lys-13, analogous to the cleavage of pig in extensively digested
purified enzyme (25). The disappearance of the doublet implies that the
modification lies extracellularly in the first 13 amino acids
(MTELSANHGGSAK).
View larger version (38K):
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Fig. 3.
The doublet results
from a post-translational modification near the N terminus.
a, right-side-out sealed renal medullary vesicles
(lane 1, control) were trypsinized at a trypsin:protein
ratio of 1:3 for 60 min at 37 °C (lane 2) (24). The doublet was reduced to a single band of lower Mr
that still stained with the antiserum against the C terminus.
b, purified enzyme from rat renal medulla was treated with 1 M Tris, pH 11 (lane 1, control), or 1 M hydroxylamine, pH 11 (lane 2), at 37 °C
overnight (23). The upper band was reduced in size to that of the
lower band by hydroxylamine, and no smaller species were
generated.
from rat, human, and mouse. Modification of Lys-13 is unlikely
because trypsin still cleaved, but modification of the N terminus was
not ruled out. The Na,K-ATPase 1 subunit in kidney is also modified
by a group that is labile to basic hydroxylamine, but this modification
is intracellular (23).
in the Kidney--
We examined the
distribution of in rat kidney by double-label confocal microscopy,
using the 1 subunit of the Na,K-ATPase for comparison because
Na,K-ATPase is known to be expressed at higher levels in some nephron
segments than in others (Fig. 4). The
anti- antibodies were raised against the C terminus and should detect forms with or without the posttranslational modification. Distal convoluted tubule (DCT) stained brightly for both 1 and ,
and proximal convoluted tubule (PCT) stained weakly for both. Other
segments, however, including cortical thick ascending limb (CTAL) and
some, but not all, cortical collecting tubules (CCT) (not shown),
expressed 1 without any detected above the background of
secondary antibody stain. Thus the expression of is
segment-specific in the rat nephron.
View larger version (39K):
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Fig. 4.
Renal cortex stained with
1- and -specific
antibodies shows nephron segments with 1 but
no . a, 1 stain, which was
brightest in distal convoluted tubule and cortical thick ascending
limb. Proximal tubule was stained, but more lightly. b, stain, which colocalized with 1 in distal convoluted tubule but not
cortical thick ascending limb, where the stain was at the level of
background. c, two-color image, scale bar 50 µm.
DCT, distal convoluted tubule; CTAL, cortical
thick ascending limb; PCT, proximal convoluted tubule.
DISCUSSION
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
expression was that the
affinities of the Na,K-ATPase for its substrates, Na+ and
K+, could be modified independently. In clones expressing
the doublet, as well as enzyme purified from renal medulla (which has
the doublet), affinities for both ions were reduced in concert. This is
not easy to explain by a shift between Na,K-ATPase E1-E2 conformations as suggested by Blostein and co-workers (19) but could be understood as
a consequence of physical association of with portions of comprising the ion binding sites. The stoichiometry of relative to
was lower than in renal medullary Na,K-ATPase, however, which raises the question of whether acts exclusively as a subunit. has been shown to form an ion channel in the absence of (14). It
has not been ruled out that the observed effects on ion affinities result from an indirect modulatory pathway activated by its presence.
modification. It is intriguing that the clones with fully modified have alteration of K+ affinity alone, while the
half-modified clones show alteration of both ion affinities. The
modification appears to nullify a major functional consequence of
association with .
is consistent with Na+
affinities measured in different nephron segments: the presence of is accompanied by lower affinity for Na+. This suggests
that expression of selectively modifies Na,K-ATPase properties
in vivo. Whether the post-translational modification demonstrated here has a specific cellular distribution among
-positive nephron segments remains to be determined.
. Regardless
of whether acts as a subunit or a channel, the evidence here
indicates that it is an important contributor to the control of renal
transport physiology.
ACKNOWLEDGEMENTS
antibodies used in
early stages of this work.
FOOTNOTES
To whom correspondence should be addressed: 149-6118;
Massachusetts General Hospital, 149 13th St., Charlestown, MA 02129. Tel.: 617-726-8579; Fax: 617-726-7526; E-mail:
sweadner@helix.mgh.harvard.edu.
ABBREVIATIONS
REFERENCES
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
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R. K. Wetzel, J. L. Pascoa, and E. Arystarkhova Stress-induced Expression of the {gamma} Subunit (FXYD2) Modulates Na,K-ATPase Activity and Cell Growth J. Biol. Chem., October 1, 2004; 279(40): 41750 - 41757. [Abstract] [Full Text] [PDF]
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C. Li, A. Grosdidier, G. Crambert, J.-D. Horisberger, O. Michielin, and K. Geering Structural and Functional Interaction Sites between Na,K-ATPase and FXYD Proteins J. Biol. Chem., September 10, 2004; 279(37): 38895 - 38902. [Abstract] [Full Text] [PDF]
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G. Crambert, C. Li, L. K. Swee, and K. Geering FXYD7, Mapping of Functional Sites Involved in Endoplasmic Reticulum Export, Association With and Regulation of Na,K-ATPase J. Biol. Chem., July 16, 2004; 279(29): 30888 - 30895. [Abstract] [Full Text] [PDF]
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A. V. Ivanov, M. E. Gable, and A. Askari Interaction of SDS with Na+/K+-ATPase: SDS-SOLUBILIZED ENZYME RETAINS PARTIAL STRUCTURE AND FUNCTION J. Biol. Chem., July 9, 2004; 279(28): 29832 - 29840. [Abstract] [Full Text] [PDF]
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 C. A. Hinojos and P. A. Doris Altered Subcellular Distribution of Na+,K+-ATPase in Proximal Tubules in Young Spontaneously Hypertensive Rats Hypertension, July 1, 2004; 44(1): 95 - 100. [Abstract] [Full Text] [PDF]
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 V. Summa, S. M. R. Camargo, C. Bauch, M. Zecevic, and F. Verrey Isoform specificity of human Na+,K+-ATPase localization and aldosterone regulation in mouse kidney cells J. Physiol., March 1, 2004; 555(2): 355 - 364. [Abstract] [Full Text] [PDF]
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J. P.T. Higgins, L. Wang, N. Kambham, K. Montgomery, V. Mason, S. U. Vogelmann, K. V. Lemley, P. O. Brown, J. D. Brooks, and M. van de Rijn Gene Expression in the Normal Adult Human Kidney Assessed by Complementary DNA Microarray Mol. Biol. Cell, February 1, 2004; 15(2): 649 - 656. [Abstract] [Full Text] [PDF]
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Y. A. Mahmmoud, G. Cramb, A. B Maunsbach, C. P. Cutler, L. Meischke, and F. Cornelius Regulation of Na,K-ATPase by PLMS, the Phospholemman-like Protein from Shark: MOLECULAR CLONING, SEQUENCE, EXPRESSION, CELLULAR DISTRIBUTION, AND FUNCTIONAL EFFECTS OF PLMS J. Biol. Chem., September 26, 2003; 278(39): 37427 - 37438. [Abstract] [Full Text] [PDF]
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A. K. Rajasekaran and S. A. Rajasekaran Role of Na-K-ATPase in the assembly of tight junctions Am J Physiol Renal Physiol, September 1, 2003; 285(3): F388 - F396. [Abstract] [Full Text] [PDF]
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R. K. Wetzel and K. J. Sweadner Phospholemman expression in extraglomerular mesangium and afferent arteriole of the juxtaglomerular apparatus Am J Physiol Renal Physiol, July 1, 2003; 285(1): F121 - F129. [Abstract] [Full Text] [PDF]
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S. A. Rajasekaran, J. Hu, J. Gopal, R. Gallemore, S. Ryazantsev, D. Bok, and A. K. Rajasekaran Na,K-ATPase inhibition alters tight junction structure and permeability in human retinal pigment epithelial cells Am J Physiol Cell Physiol, June 1, 2003; 284(6): C1497 - C1507. [Abstract] [Full Text] [PDF]
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 J. M. Capasso, C. J. Rivard, L. M. Enomoto, and T. Berl Chloride, not sodium, stimulates expression of the {gamma} subunit of Na/K-ATPase and activates JNK in response to hypertonicity in mouse IMCD3 cells PNAS, May 27, 2003; 100(11): 6428 - 6433. [Abstract] [Full Text] [PDF]
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M. Lindzen, R. Aizman, Y. Lifshitz, I. Lubarski, S. J. D. Karlish, and H. Garty Structure-Function Relations of Interactions between Na,K-ATPase, the {gamma} Subunit, and Corticosteroid Hormone-induced Factor J. Biol. Chem., May 23, 2003; 278(21): 18738 - 18743. [Abstract] [Full Text] [PDF]
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 M. S. Feschenko, C. Donnet, R. K. Wetzel, N. K. Asinovski, L. R. Jones, and K. J. Sweadner Phospholemman, a Single-Span Membrane Protein, Is an Accessory Protein of Na,K-ATPase in Cerebellum and Choroid Plexus J. Neurosci., March 15, 2003; 23(6): 2161 - 2169. [Abstract] [Full Text] [PDF]
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 I. C. Meij, L. P. W. J. van den Heuvel, S. Hemmes, W. A. van der Vliet, J. L. Willems, L. A. H. Monnens, and N. V. A. M. Knoers Exclusion of mutations in FXYD2, CLDN16 and SLC12A3 in two families with primary renal Mg2+ loss Nephrol. Dial. Transplant., March 1, 2003; 18(3): 512 - 516. [Abstract] [Full Text] [PDF]
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 G. Crambert and K. Geering FXYD Proteins: New Tissue-Specific Regulators of the Ubiquitous Na,K-ATPase Sci. Signal., January 21, 2003; 2003(166): re1 - re1. [Abstract] [Full Text] [PDF]
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H. Garty, M. Lindzen, R. Scanzano, R. Aizman, M. Fuzesi, R. Goldshleger, N. Farman, R. Blostein, and S. J. D. Karlish A functional interaction between CHIF and Na-K-ATPase: implication for regulation by FXYD proteins Am J Physiol Renal Physiol, October 1, 2002; 283(4): F607 - F615. [Abstract] [Full Text] [PDF]
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R. Aizman, C. Asher, M. Fuzesi, H. Latter, P. Lonai, S. J. D. Karlish, and H. Garty Generation and phenotypic analysis of CHIF knockout mice Am J Physiol Renal Physiol, September 1, 2002; 283(3): F569 - F577. [Abstract] [Full Text] [PDF]
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G. Crambert, M. Fuzesi, H. Garty, S. Karlish, and K. Geering Phospholemman (FXYD1) associates with Na,K-ATPase and regulates its transport properties PNAS, August 20, 2002; 99(17): 11476 - 11481. [Abstract] [Full Text] [PDF]
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H. X. Pu, R. Scanzano, and R. Blostein Distinct Regulatory Effects of the Na,K-ATPase gamma Subunit J. Biol. Chem., May 31, 2002; 277(23): 20270 - 20276. [Abstract] [Full Text] [PDF]
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J. D. Stockand New ideas about aldosterone signaling in epithelia Am J Physiol Renal Physiol, April 1, 2002; 282(4): F559 - F576. [Abstract] [Full Text] [PDF]
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E. Arystarkhova, C. Donnet, N. K. Asinovski, and K. J. Sweadner Differential Regulation of Renal Na,K-ATPase by Splice Variants of the gamma Subunit J. Biol. Chem., March 15, 2002; 277(12): 10162 - 10172. [Abstract] [Full Text] [PDF]
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G. Sweeney, W. Niu, V. A. Canfield, R. Levenson, and A. Klip Insulin increases plasma membrane content and reduces phosphorylation of Na+-K+ pump alpha 1-subunit in HEK-293 cells Am J Physiol Cell Physiol, December 1, 2001; 281(6): C1797 - C1803. [Abstract] [Full Text] [PDF]
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J. M. Capasso, C. Rivard, and T. Berl The expression of the gamma subunit of Na-K-ATPase is regulated by osmolality via C-terminal Jun kinase and phosphatidylinositol 3-kinase-dependent mechanisms PNAS, October 25, 2001; (2001) 231309198. [Abstract] [Full Text] [PDF]
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Q. Sha, K. L Lansbery, D. Distefano, R. W Mercer, and C. G Nichols Heterologous expression of the Na+,K+-ATPase {gamma} subunit in Xenopus oocytes induces an endogenous, voltage-gated large diameter pore J. Physiol., September 1, 2001; 535(2): 407 - 417. [Abstract] [Full Text] [PDF]
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R. K. Wetzel and K. J. Sweadner Immunocytochemical localization of Na-K-ATPase alpha - and gamma -subunits in rat kidney Am J Physiol Renal Physiol, September 1, 2001; 281(3): F531 - F545. [Abstract] [Full Text] [PDF]
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 D. H. JONES, M. C. GOLDING, K. J. BARR, G.-H. FONG, and G. M. KIDDER The mouse Na+-K+-ATPase {gamma}-subunit gene (Fxyd2) encodes three developmentally regulated transcripts Physiol Genomics, August 28, 2001; 6(3): 129 - 135. [Abstract] [Full Text] [PDF]
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 R. K. Wetzel and K. J. Sweadner Immunocytochemical Localization of NaK-ATPase Isoforms in the Rat and Mouse Ocular Ciliary Epithelium Invest. Ophthalmol. Vis. Sci., March 1, 2001; 42(3): 763 - 769. [Abstract] [Full Text]
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E. Feraille and A. Doucet Sodium-Potassium-Adenosinetriphosphatase-Dependent Sodium Transport in the Kidney: Hormonal Control Physiol Rev, January 1, 2001; 81(1): 345 - 418. [Abstract] [Full Text] [PDF]
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A. G. Therien and R. Blostein Mechanisms of sodium pump regulation Am J Physiol Cell Physiol, September 1, 2000; 279(3): C541 - C566. [Abstract] [Full Text] [PDF]
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B. Kuster, A. Shainskaya, H. X. Pu, R. Goldshleger, R. Blostein, M. Mann, and S. J. D. Karlish A New Variant of the gamma Subunit of Renal Na,K-ATPase. IDENTIFICATION BY MASS SPECTROMETRY, ANTIBODY BINDING, AND EXPRESSION IN CULTURED CELLS J. Biol. Chem., June 9, 2000; 275(24): 18441 - 18446. [Abstract] [Full Text] [PDF]
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A. L. Woo, P. F. James, and J. B Lingrel Sperm Motility Is Dependent on a Unique Isoform of the Na,K-ATPase J. Biol. Chem., June 30, 2000; 275(27): 20693 - 20699. [Abstract] [Full Text] [PDF]
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Y. A. Mahmmoud, H. Vorum, and F. Cornelius Identification of a Phospholemman-like Protein from Shark Rectal Glands. EVIDENCE FOR INDIRECT REGULATION OF Na,K-ATPase BY PROTEIN KINASE C VIA A NOVEL MEMBER OF THE FXYDY FAMILY J. Biol. Chem., November 10, 2000; 275(46): 35969 - 35977. [Abstract] [Full Text] [PDF]
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C. Donnet, E. Arystarkhova, and K. J. Sweadner Thermal Denaturation of the Na,K-ATPase Provides Evidence for alpha -alpha Oligomeric Interaction and gamma Subunit Association with the C-terminal Domain J. Biol. Chem., March 2, 2001; 276(10): 7357 - 7365. [Abstract] [Full Text] [PDF]
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H. X. Pu, F. Cluzeaud, R. Goldshleger, S. J. D. Karlish, N. Farman, and R. Blostein Functional Role and Immunocytochemical Localization of the gamma a and gamma b Forms of the Na,K-ATPase gamma Subunit J. Biol. Chem., June 1, 2001; 276(23): 20370 - 20378. [Abstract] [Full Text] [PDF]
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J. M. Capasso, C. Rivard, and T. Berl The expression of the gamma subunit of Na-K-ATPase is regulated by osmolality via C-terminal Jun kinase and phosphatidylinositol 3-kinase-dependent mechanisms PNAS, November 6, 2001; 98(23): 13414 - 13419. [Abstract] [Full Text] [PDF]
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S. M. Wall, M. P. Fischer, G.-H. Kim, B.-M. Nguyen, and K. A. Hassell In rat inner medullary collecting duct, NH+4 uptake by the Na,K-ATPase is increased during hypokalemia Am J Physiol Renal Physiol, January 1, 2002; 282(1): F91 - F102. [Abstract] [Full Text] [PDF]
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E. Arystarkhova, R. K. Wetzel, and K. J. Sweadner Distribution and oligomeric association of splice forms of Na+-K+-ATPase regulatory gamma -subunit in rat kidney Am J Physiol Renal Physiol, March 1, 2002; 282(3): F393 - F407. [Abstract] [Full Text] [PDF]
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