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(Received for publication, June 12, 1997)
From the Laboratory of Membrane Biology, Neuroscience Center,
Massachusetts General Hospital, Charlestown, Massachusetts 02129
The Na,K-ATPase belongs to a family of P-type
ion-translocating ATPases sharing homologous catalytic subunits (
Volume 272, Number 36,
Issue of September 5, 1997
pp. 22405-22408
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
COMMUNICATION:
3 Subunit
THE PRESENCE OF 
3 IN LUNG AND LIVER ADDRESSES THE PROBLEM OF
THE MISSING SUBUNIT
)
that traverse the membrane several times and contain the binding sites
for ATP and cations. In this family, only Na,K- and H,K-ATPases have
been shown to have a second subunit, a single-span glycoprotein called . Recently a new isoform (3) has been identified in mammals. Here
we describe structural features and tissue distribution of the 3
protein, utilizing an antiserum specific for its N terminus. 3 was
the only detected in Na,K-ATPase purified from C6 glioma. Treatment
with N-glycosidase F confirmed that 3 is a glycoprotein containing N-linked carbohydrate chains. Molecular masses
of the glycosylated protein and core protein were estimated to be 42 and 35 kDa, respectively, which are different from those of the 1
and 2 subunits. Detection of subunits has historically been difficult in certain tissues. Sensitivity was improved by
deglycosylating, and expression was evaluated by obtaining estimates of
3/ ratio. The proportion of 3 protein in the rat was highest
in lung and testis. It was also present in liver and skeletal muscle,
whereas kidney, heart, and brain contained it only as a minor component of the Na,K-ATPase. In P7 rat, we found skeletal muscle and lung Na,K-ATPase to be the most enriched in 3 subunit, whereas expression in liver was very low, illustrating developmentally regulated changes
in expression. The substantial expression in lung and adult liver very
likely explains long-standing puzzles about an apparent paucity of subunit in membranes or in discrete cellular or subcellular
structures.
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