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2.7.1.32: choline kinase

This is an abbreviated version!
For detailed information about choline kinase, go to the full flat file.

Word Map on EC 2.7.1.32

Reaction

ATP
+
choline
=
ADP
 +
phosphocholine

Synonyms

ATP/choline phosphotransferase, ATP:choline phosphotransferase, CEK1, CEK2, CEK3, chbA, CHETK-alpha, CHK, chk-alpha, Chka, CHKA2, ChKalpha, ChoK, ChoK alpha, ChoKalpha, CHOKalpha1, ChoKalpha2, ChoKbeta1, choline kinase, choline kinase alpa, choline kinase alpha, choline kinase alpha1, choline kinase alpha2, choline kinase beta, choline kinase beta1, choline kinase isoform alpha2, choline kinase-alpha, choline phosphokinase, choline-ethanolamine kinase, choline/ethanolamine kinase, choline/ethanolamine kinase 1, choline/ethanolamine kinase 2, choline/ethanolamine kinase 3, CK, CK-alpha, CK-alpha1, CK-alpha1/beta, CK-alpha2, CK-beta, CK1, CK2, CK3, CKA-2, CKalpha, CKalpha1, CKalpha2, ckb, CKbeta, CKbeta1, CKI1, CKI1-encoded choline kinase, CKII, ER-localized Cho kinase, EtnK, hChoK, kinase, choline (phosphorylating), LicA, LinfC/EK35, More, PfChoK, PfCK, protein LicA, putative choline kinase, sChoK, TbC/EK2

ECTree

     2 Transferases
         2.7 Transferring phosphorus-containing groups
             2.7.1 Phosphotransferases with an alcohol group as acceptor
                2.7.1.32 choline kinase

Cloned

Cloned on EC 2.7.1.32 - choline kinase

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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
7 different genes, cloned 5
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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.
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a series of luciferase vectors that contained the CKalpha promoter and ranged from +57 to -3840 bp is constructed. The vectors are cotransfected into hepa-1 cells with the renilla luciferase expression vector as a control
encoding isozymes alpha, beta
enzyme cloned by complementation of the yeast choline kinase mutation cki
-
enzyme expressed in Escherichia coli
-
expressed in COS-7 cells
-
expressed in Escherichia coli
expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli BL21DE3pLysS
expressed in Escherichia coli Rosetta cells and in Saccharomyces cerevisiae KS106 cells
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expressed in Escherichia coli strain BL21(DE3)
expressed in Sf9 cells
-
expression in COS-7 cells
-
gene CHKA, recombinant expression of His-SUMO-tagged wild-type and mutant enzymes in Escherichia coli strain Rosetta (DE3)pLysS
gene CK1, recombinant overexpression in Arabidopsis thaliana plants via Agrobacterium tumefaciens-mediated transformation, quantitative RT-PCR enzyme expression analysis at normal or tunicamycin (Tm)-induced endoplasmic reticulum (ER) stress conditions
gene licA, recombinant expression of enzyme sChoK in Escherichia coli strain BL21(DE3)
gene licA, sequence comparison of Streptococcus pneumoniae and human enzymes, overview
His-tag, expressed in Escherichia coli BL21(DE3)-pRIL
locus At1g74320, recombinant overexpression in Arabidopsis thaliana plants via Agrobacterium tumefaciens-mediated transformation, quantitative RT-PCR enzyme expression analysis at normal or tunicamycin (Tm)-induced endoplasmic reticulum (ER) stress conditions
locus At4g09760, recombinant overexpression in Arabidopsis thaliana plants via Agrobacterium tumefaciens-mediated transformation, quantitative RT-PCR enzyme expression analysis at normal or tunicamycin (Tm)-induced endoplasmic reticulum (ER) stress conditions
molecular cloning and recombinant expression in Escherichia coli strains DH5alpha, XL-1 blue and TOP10. BL21-Gold(DE3) is used for protein overexpression, RNA from bloodstream form of Trypanosoma brucei is used for cDNA synthesis.
-
native and mutant enzymes expressed as His-tag fusion protein in Sf9 cells
-
overview
parts of the alpha and beta subtype expressed as GST-fusion protein in Escherichia coli, alpha and beta subtype coexpressed in COS-7 cells
-
plasmid maintenance and amplification are performed in Escherichia coli strain DH5 alpha
-
recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3) Star
recombinant expression of tagged choline kinase alpha1 residues 75-457
Small interfering RNA-chk (siRNA) is designed and transfected for 48 h using oligofectamine and Opti-MEM based on the observed decrease of Chk message using reverse transcription-PCR analysis within this time. Immunoblot assay results with Chk antibody shows significant reduction of Chk protein expression levels in MCF-12A, MCF-7, and MDA-MB-231 cells after transient siRNA-chk transfection.
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The genomic sequence of the putative human chk-alpha promoter is obtained by performing a BLAST search in National Center for Biotechnology Information using the known promoter sequence of the rat chk-a gene as query sequence. A 2.3-kb region at the 5'-end of the human chk-a gene on chromosome 11, immediately upstream of the translation start site, is found to contain several putative HRE sites with the core sequence RCGTG. Due to its high GC content, this 2.3-kb region is cloned as four smaller overlapping fragments, and formamide (5%) is added in all PCRs. The four overlapping sequences are PCR amplified from human genomic DNA. The PCR products of paired sense and antisense primer sequences used for the cloning of these fragments (Chk) are subcloned into the pCR 2.1 TA vector. The full 2.3-kb promoter region is obtained by appropriate endonuclease digestions and ligation of these four overlapping fragments. Each of the four fragments alone, as well as several subligations of two or three consecutive fragments, and the full-length 2.3-kb chk-a promoter are cloned into the pGL4-basic Luc reporter vector.
-
The RT-PCR-amplified PfCK gene is cloned in pRSET-C, an Escherichia coli expression vector. The host strain is optimized
-
To identify the region of the CK apha promoter that is important to regulate its transcription, a series of luciferase vectors that contained the CK alpha promoter and ranged from +57 to -3840 bp are constructed. The vectors are cotransfected into Hepa-1 cells with the renilla luciferase expression vector as a control
Transfection of human HeK293T is carried out by the calcium phosphate method. The amount of plasmidic DNA is kept constant at 0.2 mg with the corresponding empty vector. Tumors induced by ChoK alpha over-expression in Hek293T are removed from the mice to generate the ChoK tumoral cell line ADJ. Different slices of the tissue are placed into culture 100mm dishes and treated with trypsin and G418 to avoid murine cells contamination. The resultant cell line (ADJ) displays constitutive ChoK alpha activation and a fully transformed phenotype
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