Impaired plasma membrane targeting or protein stability by certain ATP1A2 mutations identified in sporadic or familial hemiplegic migraine.
Mutations in three different genes have been implicated in familial hemiplegic migraine (FHM), two of them code for neuronal voltage-gated cation channels, CACNA1A and SCN1A, while the third encodes ATP1A2, the alpha(2)-isoform of the Na(+)/K(+)-ATPase's catalytic subunit, thus classifying FHM as an ion channel/ion transporter disorder. The Na(+)/K(+)-ATPase maintains the physiological gradients for Na(+) and K(+) ions and is therefore critical for the activity of ion channels and transporters involved in neurotransmitter uptake or Ca(2+) signaling. Diverse functional abnormalities have been identified for disease-linked ATP1A2 mutations, which reach far beyond simple loss-of-function. We have shown recently that ATP1A2 mutations frequently lead to changes in the enzyme's voltage-dependent properties, kinetics or apparent cation affinities. Here, we present functional data on a so far uncharacterized set of ATP1A2 mutations (G301R, R908Q and P979L) upon expression in Xenopus oocytes and HEK293FT cells, and provide evidence for a novel pathophysiological mechanism. Whereas the G301R mutant was inactive, no functional changes were observed for mutants R908Q and P979L in the oocyte expression system. However, the R908Q mutant was less effectively expressed in the plasma membrane of oocytes, making it the first missense mutation to result in defective plasma membrane targeting. Notably, the P979L mutant exhibited the same cellular expression profile as the wild-type protein, both in Xenopus oocytes and in transfected HEK293FT cells grown at 28 degrees C, but much less P979L protein was found upon cell growth at 37 degrees C, showing for the first time that temperature-sensitive effects on protein stability can underlie ATP1A2 loss-of-function.
Technical University of Berlin, Institute of Chemistry, Berlin, Germany.
Neslihan N
NN
Tavraz
Katharina L
KL
Dürr
Jan B
JB
Koenderink
Tobias
T
Freilinger
Ernst
E
Bamberg
Martin
M
Dichgans
Thomas
T
Friedrich
D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D006801:Humans; D008881:Migraine Disorders / Q000235:genetics; D020325:Migraine with Aura / Q000235:genetics*; D020125:Mutation, Missense; D009865:Oocytes; D055550:Protein Stability; D021381:Protein Transport; D000254:Sodium-Potassium-Exchanging ATPase / Q000235:genetics* / Q000378:metabolism*; D013696:Temperature; D014161:Transduction, Genetic; D014981:Xenopus
C498531:ATP1A2 protein, human; D000254:Sodium-Potassium-Exchanging ATPase