Literature summary for 2.7.1.32 extracted from

Aoyama, C.; Young, S.G.; Vance, D.E.
Early embryonic lethality caused by disruption of the gene for choline kinase alpha, the first enzyme in phosphatidylcholine biosynthesis (2008), J. Biol. Chem., 283, 1456-1462.

Cloned(Commentary)

Cloned (Comment)	Organism
A mouse embryonic stem cell line (XH252, strain 129/OlaHsd) with an insertional mutation in choline kinase alpha is created in a gene-trapping prgram. The vector, pGT1Lxf, is designed to create an in-frame fusion between the 5' exons of the trapped gene and a reporter, betageo (a fusion of beta-galactosidase and neomycin phosphotransferase 2). CK-alpha spans 12 exons on mouse chromosome 19. The insertional mutation in XH252 occurred in intron 5. Thus, the gene-trapped locus is predicted to yield a fusion transcript containing exons 1-5 of CK-alpha and betageo. The embryonic stem cells are injected into C57BL/6 blastocysts to create chimeric mice, which are bred with C57BL/6 mice to generate heterozygous (+/-) CK-alpha-deficient mice. All mice had a mixed genetic background. The mice are weaned at 21 days of age, housed in a barrier facility with a 12-h light-dark cycle, and fed chow containing 4.5% fat. Creating of mice lacking CK- alpha with an embryonic stem cell line containing an insertional mutation in the gene for CK-alpha. Embryos homozygous for the mutant CK-alpha allele are recovered at the blastocyst stage, but not at embryonic day 7.5, indicating that CK-alpha is crucial for the early development of mouse embryos.	Mus musculus

Cloned (Comment)

Organism

A mouse embryonic stem cell line (XH252, strain 129/OlaHsd) with an insertional mutation in choline kinase alpha is created in a gene-trapping prgram. The vector, pGT1Lxf, is designed to create an in-frame fusion between the 5' exons of the trapped gene and a reporter, betageo (a fusion of beta-galactosidase and neomycin phosphotransferase 2). CK-alpha spans 12 exons on mouse chromosome 19. The insertional mutation in XH252 occurred in intron 5. Thus, the gene-trapped locus is predicted to yield a fusion transcript containing exons 1-5 of CK-alpha and betageo. The embryonic stem cells are injected into C57BL/6 blastocysts to create chimeric mice, which are bred with C57BL/6 mice to generate heterozygous (+/-) CK-alpha-deficient mice. All mice had a mixed genetic background. The mice are weaned at 21 days of age, housed in a barrier facility with a 12-h light-dark cycle, and fed chow containing 4.5% fat. Creating of mice lacking CK- alpha with an embryonic stem cell line containing an insertional mutation in the gene for CK-alpha. Embryos homozygous for the mutant CK-alpha allele are recovered at the blastocyst stage, but not at embryonic day 7.5, indicating that CK-alpha is crucial for the early development of mouse embryos.

Mus musculus

Inhibitors

Inhibitors	Comment	Organism	Structure
additional information	the lethality of the homozygous choline kinase alpha knock-out mice embryos indicates the indispensable role of CK-alphain early embryogenesis	Mus musculus

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
cytosol	assay at	Mus musculus	5829	-

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
choline + ATP	Mus musculus	catalyzes the first phosphorylation reaction in the Kennedy pathway (biosynthesis of the major membrane phospholipid phosphatidylcholine)	phosphocholine + ADP	-	?

Organism

Organism	UniProt	Comment	Textmining
Mus musculus	-	knock-out mice, heterozygous, +/-choline kinase alpha deficient	-

Source Tissue

Source Tissue	Comment	Organism	Textmining
liver	-	Mus musculus	-
skeletal muscle	forelimb, hindlimb	Mus musculus	-
testis	-	Mus musculus	-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.000139	-	skeletal muscle, hindlimbs of +/+ CK-alpha mice, supernatant of tissue homogenates are incubated in volume of 0.100 ml of reaction buffer at 37°C for 30 min. The reaction product, phosphocholine, is separated using an AG1-X8 column.To determine the activity of each CK isoform, supernatant fractions are treated with an antisera raised against GST (control), GST-CK-alpha or GST-CK-beta fusion proteins combined with protein A-Sepharose overnight at 4°C, and the supernatant is used.	Mus musculus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
choline + ATP	catalyzes the first phosphorylation reaction in the Kennedy pathway (biosynthesis of the major membrane phospholipid phosphatidylcholine)	Mus musculus	phosphocholine + ADP	-	?

Synonyms

Synonyms	Comment	Organism
choline kinase alpha	-	Mus musculus
CK	-	Mus musculus
CK-alpha	-	Mus musculus

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Mus musculus

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8.8	-	assay at	Mus musculus