Literature summary for 7.2.2.10 extracted from

Tidow, H.; Hein, K.L.; Baekgaard, L.; Palmgren, M.G.; Nissen, P.
Expression, purification, crystallization and preliminary X-ray analysis of calmodulin in complex with the regulatory domain of the plasma-membrane Ca2+-ATPase ACA8 (2010), Acta Crystallogr. Sect. F, 66, 361-363.

Activating Compound

Activating Compound	Comment	Organism	Structure
calcium-bound calmodulin	in the resting state, the plant plasma-membrane Ca2+-ATPase is autoinhibited by binding of its N-terminal tail to two major intracellular loops. Activation requires the binding of calcium-bound calmodulin to this tail and a conformational change that displaces the autoinhibitory tail from the catalytic domain	Arabidopsis thaliana

Cloned(Commentary)

Cloned (Comment)	Organism
expression of ACA8 residues 40-95, comprising the calmodulin binding site of the enzyme, in Escherichia coli as protein with an N-terminal fusion consisting of a His6 tag, a lipoyl domain and a TEV protease cleavage site	Arabidopsis thaliana

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant complex between calmodulin and the regulatory domain of the plasma-membrane Ca2+-ATPase ACA8, mixing of 0.0006 ml of protein solution, containing 16 mg/ml protein in 25 mM Tris, pH 7.0, 50 mM NaCl, 10 mM 2-mercaptoethanol, 5 mM CaCl2, with 0.001 ml reservoir solution, containing 2.0 M ammonium sulfate, 0.1 M CAPS, pH 10.5, 0.2 M lithium sulfate, at final pH 8.2, ar 20°C, X-ray diffraction structure determination and analysis at 3.0 A resolution	Arabidopsis thaliana

Crystallization (Comment)

Organism

purified recombinant complex between calmodulin and the regulatory domain of the plasma-membrane Ca2+-ATPase ACA8, mixing of 0.0006 ml of protein solution, containing 16 mg/ml protein in 25 mM Tris, pH 7.0, 50 mM NaCl, 10 mM 2-mercaptoethanol, 5 mM CaCl2, with 0.001 ml reservoir solution, containing 2.0 M ammonium sulfate, 0.1 M CAPS, pH 10.5, 0.2 M lithium sulfate, at final pH 8.2, ar 20°C, X-ray diffraction structure determination and analysis at 3.0 A resolution

Arabidopsis thaliana

Inhibitors

Inhibitors	Comment	Organism	Structure
additional information	in the resting state, the plant plasma-membrane Ca2+-ATPase is autoinhibited by binding of its N-terminal tail to two major intracellular loops. Activation requires the binding of calcium-bound calmodulin to this tail and a conformational change that displaces the autoinhibitory tail from the catalytic domain	Arabidopsis thaliana

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
plasma membrane	-	Arabidopsis thaliana	5886	-

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Ca2+	in the resting state, the plant plasma-membrane Ca2+-ATPase is autoinhibited by binding of its N-terminal tail to two major intracellular loops. Activation requires the binding of calcium-bound calmodulin to this tail and a conformational change that displaces the autoinhibitory tail from the catalytic domain	Arabidopsis thaliana

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + H2O + Ca2+/in	Arabidopsis thaliana	plasma-membrane Ca2+-ATPase is a calcium pump that exports Ca2+ from the cytosol to the extracellular environment of eukaryotic cells and thus maintain overall Ca2+ homoeostasis and provide local control of intracellular Ca2+ signalling	ADP + phosphate + Ca2+/out	-	?

Organism

Organism	UniProt	Comment	Textmining
Arabidopsis thaliana	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His6-tagged ACA8 residues 40-95, comprising the calmodulin binding site of the enzyme, from Escherichia coli by nickel affinity chromatography and gel filtration to homogeneity	Arabidopsis thaliana

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + H2O + Ca2+/in	-	Arabidopsis thaliana	ADP + phosphate + Ca2+/out	-	?
ATP + H2O + Ca2+/in	plasma-membrane Ca2+-ATPase is a calcium pump that exports Ca2+ from the cytosol to the extracellular environment of eukaryotic cells and thus maintain overall Ca2+ homoeostasis and provide local control of intracellular Ca2+ signalling	Arabidopsis thaliana	ADP + phosphate + Ca2+/out	-	?

Synonyms

Synonyms	Comment	Organism
ACA	-	Arabidopsis thaliana
ACA8	-	Arabidopsis thaliana
autoinhibited Ca2+-ATPase	-	Arabidopsis thaliana
More	the enzyme belongs to the P2B subfamily of P-type ATPases	Arabidopsis thaliana
plasma-membrane Ca2+-ATPase	-	Arabidopsis thaliana
PMCA	-	Arabidopsis thaliana

General Information

General Information	Comment	Organism
physiological function	plasma-membrane Ca2+-ATPases, PMCAs, are high-affinity calcium pumps that expel Ca2+ from eukaryotic cells to maintain overall Ca2+ homoeostasis and to provide local control of intracellular Ca2+ signalling. They are of major physiological importance, with different isoforms being essential, for example, for presynaptic and postsynaptic Ca2+ regulation in neurons, feedback signalling in the heart and sperm motility	Arabidopsis thaliana