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The Journal of Neuroscience, March 15, 2003, 23(6):2161
Phospholemman, a Single-Span Membrane Protein, Is an Accessory
Protein of Na,K-ATPase in Cerebellum and Choroid Plexus
Marina S.
Feschenko1,
Claudia
Donnet1,
Randall K.
Wetzel1,
Natalya K.
Asinovski1,
Larry R.
Jones2, and
Kathleen J.
Sweadner1
1 Laboratory of Membrane Biology, Neuroscience Center,
Massachusetts General Hospital, Charlestown, Massachusetts 02129, and
2 Krannert Institute of Cardiology, Indiana University
School of Medicine, Indianapolis, Indiana 46202
Phospholemman (FXYD1) is a homolog of the Na,K-ATPase  subunit
(FXYD2), a small accessory protein that modulates ATPase activity. Here
we show that phospholemman is highly expressed in selected structures
in the CNS. It is most abundant in cerebellum, where it was detected in
the molecular layer, in Purkinje neurons, and in axons traversing the
granule cell layer. Phospholemman was particularly enriched in choroid
plexus, the organ that secretes CSF in the ventricles, where it
colocalized with Na,K-ATPase in the apical membrane. It was also
enriched, with Na,K-ATPase, in certain tanycytes or ependymal cells of
the ventricle wall. Two different experimental approaches demonstrated
that phospholemman physically associated with the Na,K-ATPase in
cerebellum and choroid plexus: the proteins copurified after detergent
treatment and co-immunoprecipitated from solubilized crude membranes
using either anti-phospholemman or anti-Na,K-ATPase antibodies.
Phospholemman antibodies precipitated all three Na,K-ATPase  subunit
isoforms (1-3) from cerebellum, indicating that the interaction
is not specific to a particular isoform and consistent with the
presence of phospholemman in both neurons and glia. Antibodies against the C-terminal domain of phospholemman reduced Na,K-ATPase activity in vitro without effect on Na+
affinity. At least two other FXYD family members have been detected in
the CNS, suggesting that additional complexity of sodium pump regulation will be found.
Key words:
Na,K-ATPase; phospholemman; FXYD; cerebellum; choroid plexus; Purkinje cell; Bergmann glia
Copyright © 2003 Society for Neuroscience 0270-6474/03/2362161-09$05.00/0
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