Phospholipids and calmodulin modulate the inhibition of PMCA activity by tau.
The disruption of Ca2+signaling in neurons, together with a failure to keep optimal intracellular Ca2+concentrations, have been proposed as significant factors for neuronal dysfunction in the Ca2+hypothesis of Alzheimer's disease (AD). Tau is a protein that plays an essential role in axonal transport and can form abnormal structures such as neurofibrillary tangles that constitute one of the hallmarks of AD. We have recently shown that plasma membrane Ca2+-ATPase (PMCA), a key enzyme in the maintenance of optimal cytosolic Ca2+levels in cells, is inhibited by tau in membrane vesicles. In the present study we show that tau inhibits synaptosomal PMCA purified from pig cerebrum, and reconstituted in phosphatidylserine-containing lipid bilayers, with a Kivalue of 1.5±0.2nM tau. Noteworthy, the inhibitory effect of tau is dependent on the charge of the phospholipid used for PMCA reconstitution. In addition, nanomolar concentrations of calmodulin, the major endogenous activator of PMCA, protects against inhibition of the Ca2+-ATPase activity by tau. Our results in a cellular model such as SH-SY5Y human neuroblastoma cells yielded an inhibition of PMCA by nanomolar tau concentrations and protection by calmodulin against this inhibition similar to those obtained with purified synaptosomal PMCA. Functional studies were also performed with native and truncated versions of human cerebral PMCA4b, an isoform that has been showed to be functionally regulated by amyloid peptides, whose aggregates constitutes another hallmark of AD. Kinetic assays point out that tau binds to the C-terminal tail of PMCA, at a site distinct but close to the calmodulin binding domain. In conclusion, PMCA can be seen as a molecular target for tau-induced cytosolic calcium dysregulation in synaptic terminals. This article is part of a Special Issue entitled: ECS Meeting edited by Claus Heizmann, Joachim Krebs and Jacques Haiech.
Departamento de Bioquímica y Biología Molecular y Genética, Facultad de Ciencias, Universidad de Extremadura, 06006 Badajoz, Spain.
María
M
Berrocal
Departamento de Bioquímica y Biología Molecular y Genética, Facultad de Ciencias, Universidad de Extremadura, 06006 Badajoz, Spain.
Isaac
I
Corbacho
Departamento de Bioquímica y Biología Molecular y Genética, Facultad de Ciencias, Universidad de Extremadura, 06006 Badajoz, Spain.
M Rosario
MR
Sepulveda
Departamento de Bioquímica y Biología Molecular y Genética, Facultad de Ciencias, Universidad de Extremadura, 06006 Badajoz, Spain.
Carlos
C
Gutierrez-Merino
Departamento de Bioquímica y Biología Molecular y Genética, Facultad de Ciencias, Universidad de Extremadura, 06006 Badajoz, Spain. Electronic address: anam@unex.es.
Ana M
AM
Mata
D000818:Animals; D002147:Calmodulin / Q000378:metabolism*; D045744:Cell Line, Tumor; D006801:Humans; D010743:Phospholipids / Q000378:metabolism*; D053499:Plasma Membrane Calcium-Transporting ATPases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D013552:Swine; D016875:tau Proteins / Q000378:metabolism*
D002147:Calmodulin; D010743:Phospholipids; D016875:tau Proteins; D053499:Plasma Membrane Calcium-Transporting ATPases
Ca(2+); Calmodulin; PMCA; Phospholipid; Tau