2.7.1.30: glycerol kinase
This is an abbreviated version!
For detailed information about glycerol kinase, go to the full flat file.
Word Map on EC 2.7.1.30
-
2.7.1.30
-
glycerol-3-phosphate
-
triglyceride
-
adrenal
-
hypoplasia
-
dystrophy
-
muscular
-
adipose
-
3-phosphate
-
duchenne
-
lipase
-
phosphoenolpyruvate
-
hexokinase
-
x-linked
-
dihydroxyacetone
-
contiguous
-
adipocytes
-
triacylglycerols
-
carboxykinase
-
congenita
-
gluconeogenesis
-
glycerophosphate
-
lipolysis
-
hpr
-
iiaglc
-
1,6-bisphosphate
-
hypogonadism
-
co-immobilized
-
phosphocarrier
-
glyceroneogenesis
-
hypogonadotropic
-
glycosomes
-
dissimilation
-
triolein
-
1,3-propanediol
-
glucose-specific
-
aquaglyceroporins
-
d-glyceraldehyde
-
phosphoenolpyruvate:sugar
-
dhap
-
phosphoenolpyruvate-dependent
-
sn-glycerol-3-phosphate
-
sn-glycerol
-
drug development
-
diagnostics
-
synthesis
- 2.7.1.30
- glycerol-3-phosphate
- triglyceride
- adrenal
- hypoplasia
- dystrophy
- muscular
- adipose
- 3-phosphate
-
duchenne
- lipase
- phosphoenolpyruvate
- hexokinase
-
x-linked
- dihydroxyacetone
-
contiguous
- adipocytes
- triacylglycerols
-
carboxykinase
- congenita
-
gluconeogenesis
- glycerophosphate
-
lipolysis
- hpr
- iiaglc
- 1,6-bisphosphate
- hypogonadism
-
co-immobilized
-
phosphocarrier
-
glyceroneogenesis
-
hypogonadotropic
- glycosomes
-
dissimilation
- triolein
- 1,3-propanediol
-
glucose-specific
-
aquaglyceroporins
- d-glyceraldehyde
-
phosphoenolpyruvate:sugar
- dhap
-
phosphoenolpyruvate-dependent
- sn-glycerol-3-phosphate
- sn-glycerol
- drug development
- diagnostics
- synthesis
Reaction
Synonyms
AFUB_068560, ASTP
ECTree
2.7.1.30 glycerol kinaseAdvanced search results
results (
results (Engineering
Engineering on EC 2.7.1.30 - glycerol kinase
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
top
PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
A344V
the mutant shows a higher melting temperature than the wild type enzyme
A344V/T386I/F388Y
the mutant with decreased catalytic efficiency shows a higher melting temperature than the wild type enzyme
C292A
the mutant shows a lower melting temperature than the wild type enzyme
L274M
the mutant shows a higher melting temperature than the wild type enzyme
L274M/T386I/F388Y
the mutant shows a higher melting temperature than the wild type enzyme
Q50E/T386I/F388Y
the mutant shows a higher melting temperature than the wild type enzyme
T386I
the mutant shows a higher melting temperature than the wild type enzyme
T386I/F388Y
the mutant with decreased catalytic efficiency shows a 9°C higher melting temperature than the wild type enzyme
A344V
-
the mutant shows a higher melting temperature than the wild type enzyme
-
C292A
-
the mutant shows a lower melting temperature than the wild type enzyme
-
L274M
-
the mutant shows a higher melting temperature than the wild type enzyme
-
T386I
-
the mutant shows a higher melting temperature than the wild type enzyme
-
T386I/F388Y
-
the mutant with decreased catalytic efficiency shows a 9°C higher melting temperature than the wild type enzyme
-
S329D
H232A
H232R
-
residue located in the activation loop, mutant protein has enhanced activity
A65T
D72V
-
the catalytic properties of the mutant differ little from those of the wild type enzyme. The mutant shows 14.76% expression compared to the wild type enzyme
E478C
-
mutation increases the affinity for glucose-specific phosphocarrier protein of the phosphoenolpyruvate:glucose phosphotransferase system (IIA(Glc))
E478C/T428V/R429N
-
T428V and R429N replace two coupling locus amino acids with those from Haemophilus influenzae glycerol kinase
G230D
G427D/T428V/R429N
-
replacement of all three of the coupling locus amino acids with those from Haemophilus influenzae glycerol kinase
I474A
the maximum extent of IIAGlc inhibition is reduced for the mutant enzyme
I474C
the maximum extent of IIAGlc inhibition is reduced for the mutant enzyme
M271I
-
the mutant shows strongly increased Km for ATP and 30.75% expression compared to the wild type enzyme
Q37P
-
the mutant shows strongly increased Km for ATP and 65.73% expression compared to the wild type enzyme
R369A
oligomeric interactions are disturbed by the amino acid substitution
R479A
the maximum extent of IIAGlc inhibition is reduced for the mutant enzyme
R479C
the maximum extent of IIAGlc inhibition is reduced for the mutant enzyme
V61L
-
the catalytic properties of the mutant differ little from those of the wild type enzyme. The mutant shows 12.71% expression compared to the wild type enzyme
E398D
naturally occurring mutation in patients with glyceroluria, causes a strong decrease in enzyme activity
G280A
-
naturally occurring mutation in a patient with glyceroluria, causes a strong decrease in enzyme activity, mutation affects a highly conserved amino acid in the ATP-binding domain
L61P
naturally occurring mutation in patients with glyceroluria, causes an 5-10-fold increased Km for glycerol
K271E
E478A
about 25% activity compared to the wild type enzyme
T12V
less than 5% activity compared to the wild type enzyme
T273V
about 80% activity compared to the wild type enzyme
additional information
H232A
-
lacks the site of activation by phosphorylation, activity similar to unphosphorylated native enzyme
A65T
oligomeric interactions are disturbed by the amino acid substitution
G230D
structural analysis reveal that the decreased allosteric regulation in the G230D mutant is a result of the altered fructose 1,6-bisphosphate binding loop conformations in the mutant that interfere with the wild-type fructose 1,6-bisphosphate binding site. The altered fructose 1,6-bisphosphate binding loop conformation in the G230D mutant of glycerol kinase are supported through a series of intramolecular loop interactions. The appearence of Asp230 in the fructose 1,6-bisphosphate binding loops also repositions the wildtype fructose 1,6-bisphosphate binding residues away from the fructose 1,6-bisphosphate binging site.
K271E
site-directed mutagenesis, the mutation disrupts the hexamer formation interface
K271E
-
site-directed mutagenesis, the mutation disrupts the hexamer formation interface
-
K271E
-
site-directed mutagenesis, the mutation disrupts the hexamer formation interface
-
additional information
knockout glcA in the DELTAgfdA and the parental wild-type strain backgrounds. The glcA null mutant shows defects of growth and conidiation in glycerol media but not in glucose media, suggesting glcA is required when glycerol is the sole carbon source, and gfdA is required when glucose is the sole carbon source. The DELTAgfdADELTAglcA double mutant shows an exacerbation of colony defects in both glucose and glycerol media. In glucose medium, DELTAgfdADELTAglcA colonies are very small, and colonies are nearly undetectable colonies in the glycerol medium. Transformation of the full-length ORF sequence of glcA under the control of the constitutive promoter gpdA into the DELTAgfdA and reference strains, resulting in two glcA-overexpression strains, DELTAgfdAOE::glcA and wild-type OE::glcA. Overexpressed glcA is able to rescue growth defects associated with loss of gfdA. In comparison, wild-type OE::glcA still shows wild-type like colony phenotypes. Overexpression of glcA may result in the production of accumulated glycerol 3-phosphate (G3P), which allows colonies to bypass the requirement of gfdA to produce glycerol use in colony growth, indicating that the growth defects of gfdA null mutant might be due to absence of G3P rather than glycerol. In contrast, when glycerol is used as the sole carbon source, DELTAgfdAOE::glcA shows defective phenotypes similar to wild-type OE::glcA, indicating the overexpression of glcA may cause the production of accumulated G3P, which results in growth defects in glycerol media either in the presence or absence of gfdA
additional information
-
knockout glcA in the DELTAgfdA and the parental wild-type strain backgrounds. The glcA null mutant shows defects of growth and conidiation in glycerol media but not in glucose media, suggesting glcA is required when glycerol is the sole carbon source, and gfdA is required when glucose is the sole carbon source. The DELTAgfdADELTAglcA double mutant shows an exacerbation of colony defects in both glucose and glycerol media. In glucose medium, DELTAgfdADELTAglcA colonies are very small, and colonies are nearly undetectable colonies in the glycerol medium. Transformation of the full-length ORF sequence of glcA under the control of the constitutive promoter gpdA into the DELTAgfdA and reference strains, resulting in two glcA-overexpression strains, DELTAgfdAOE::glcA and wild-type OE::glcA. Overexpressed glcA is able to rescue growth defects associated with loss of gfdA. In comparison, wild-type OE::glcA still shows wild-type like colony phenotypes. Overexpression of glcA may result in the production of accumulated glycerol 3-phosphate (G3P), which allows colonies to bypass the requirement of gfdA to produce glycerol use in colony growth, indicating that the growth defects of gfdA null mutant might be due to absence of G3P rather than glycerol. In contrast, when glycerol is used as the sole carbon source, DELTAgfdAOE::glcA shows defective phenotypes similar to wild-type OE::glcA, indicating the overexpression of glcA may cause the production of accumulated G3P, which results in growth defects in glycerol media either in the presence or absence of gfdA
additional information
-
knockout glcA in the DELTAgfdA and the parental wild-type strain backgrounds. The glcA null mutant shows defects of growth and conidiation in glycerol media but not in glucose media, suggesting glcA is required when glycerol is the sole carbon source, and gfdA is required when glucose is the sole carbon source. The DELTAgfdADELTAglcA double mutant shows an exacerbation of colony defects in both glucose and glycerol media. In glucose medium, DELTAgfdADELTAglcA colonies are very small, and colonies are nearly undetectable colonies in the glycerol medium. Transformation of the full-length ORF sequence of glcA under the control of the constitutive promoter gpdA into the DELTAgfdA and reference strains, resulting in two glcA-overexpression strains, DELTAgfdAOE::glcA and wild-type OE::glcA. Overexpressed glcA is able to rescue growth defects associated with loss of gfdA. In comparison, wild-type OE::glcA still shows wild-type like colony phenotypes. Overexpression of glcA may result in the production of accumulated glycerol 3-phosphate (G3P), which allows colonies to bypass the requirement of gfdA to produce glycerol use in colony growth, indicating that the growth defects of gfdA null mutant might be due to absence of G3P rather than glycerol. In contrast, when glycerol is used as the sole carbon source, DELTAgfdAOE::glcA shows defective phenotypes similar to wild-type OE::glcA, indicating the overexpression of glcA may cause the production of accumulated G3P, which results in growth defects in glycerol media either in the presence or absence of gfdA
-
additional information
-
knockout glcA in the DELTAgfdA and the parental wild-type strain backgrounds. The glcA null mutant shows defects of growth and conidiation in glycerol media but not in glucose media, suggesting glcA is required when glycerol is the sole carbon source, and gfdA is required when glucose is the sole carbon source. The DELTAgfdADELTAglcA double mutant shows an exacerbation of colony defects in both glucose and glycerol media. In glucose medium, DELTAgfdADELTAglcA colonies are very small, and colonies are nearly undetectable colonies in the glycerol medium. Transformation of the full-length ORF sequence of glcA under the control of the constitutive promoter gpdA into the DELTAgfdA and reference strains, resulting in two glcA-overexpression strains, DELTAgfdAOE::glcA and wild-type OE::glcA. Overexpressed glcA is able to rescue growth defects associated with loss of gfdA. In comparison, wild-type OE::glcA still shows wild-type like colony phenotypes. Overexpression of glcA may result in the production of accumulated glycerol 3-phosphate (G3P), which allows colonies to bypass the requirement of gfdA to produce glycerol use in colony growth, indicating that the growth defects of gfdA null mutant might be due to absence of G3P rather than glycerol. In contrast, when glycerol is used as the sole carbon source, DELTAgfdAOE::glcA shows defective phenotypes similar to wild-type OE::glcA, indicating the overexpression of glcA may cause the production of accumulated G3P, which results in growth defects in glycerol media either in the presence or absence of gfdA
-
additional information
Aspergillus fumigatus FGSC A1163
-
knockout glcA in the DELTAgfdA and the parental wild-type strain backgrounds. The glcA null mutant shows defects of growth and conidiation in glycerol media but not in glucose media, suggesting glcA is required when glycerol is the sole carbon source, and gfdA is required when glucose is the sole carbon source. The DELTAgfdADELTAglcA double mutant shows an exacerbation of colony defects in both glucose and glycerol media. In glucose medium, DELTAgfdADELTAglcA colonies are very small, and colonies are nearly undetectable colonies in the glycerol medium. Transformation of the full-length ORF sequence of glcA under the control of the constitutive promoter gpdA into the DELTAgfdA and reference strains, resulting in two glcA-overexpression strains, DELTAgfdAOE::glcA and wild-type OE::glcA. Overexpressed glcA is able to rescue growth defects associated with loss of gfdA. In comparison, wild-type OE::glcA still shows wild-type like colony phenotypes. Overexpression of glcA may result in the production of accumulated glycerol 3-phosphate (G3P), which allows colonies to bypass the requirement of gfdA to produce glycerol use in colony growth, indicating that the growth defects of gfdA null mutant might be due to absence of G3P rather than glycerol. In contrast, when glycerol is used as the sole carbon source, DELTAgfdAOE::glcA shows defective phenotypes similar to wild-type OE::glcA, indicating the overexpression of glcA may cause the production of accumulated G3P, which results in growth defects in glycerol media either in the presence or absence of gfdA
-
additional information
-
co-immobilization of nanoparticles of lipase, glycerol kinase and glycerol 3-phosphate oxidase onto pencil graphite electrode, function as improved amperometric triglyceride biosensor. An improved amperometric triglyceride (TG) biosensor is fabricated using lipaseNPs/GKNPs/GPONPs/PG electrode as the working electrode, Ag/AgCl as the standard electrode and Pt wire as auxiliary electrode. The biosensor shows optimum response within 2.5 s at pH 7.0 and temperature of 35°C. The biosensor measures current due to electrons generated at 0.1 V against Ag/AgCl, from H2O2, which is produced from triolein by co-immobilized enzyme nanoparticles (ENPs). The biosensor is evaluated and employed for determination of triglyceride in the serum of apparently healthy subject and persons suffering from hypertriglyceridemia. The biosensor loses 20% of its initial activity after continued uses over a period of 240 days, while being stored at 4°C. Construction of the biosensor, evaluation, and optimization, overview
additional information
-
rational design of nanoparticle platforms for cutting-the-fat, covalent immobilization of lipase, glycerol kinase, and glycerol-3-phosphate oxidase on metal nanoparticles. Co-immobilization of enzymes onto nanoparticles aggregates is expected to produce faster kinetics than their individual immobilizations on separate matrices. The combined activities of co-immobilized enzymes are tested amperometrically, and these composite nanobiocatalysts show optimum activity within 4-5 s, at pH 6.5-7.5 and 35°C, when polarized at a potential between 0.1 and 0.4 V. Co-immobilized enzymes show excellent linearity within 50-700 mg/dl of the lipid with detection limit of 20 mg/dl for triolein. The half life of co-immobilized enzymes is 7 months, when stored dry at 4°C, which is very convenient for practical applications. Co-immobilized biocatalysts measured triglycerides (TGs) in the sera of apparently healthy persons and persons suffering from hypertriglyceridemia, which is recognized as a leading cause for heart disease. The measurement of serum TG by co-immobilized enzymes is unaffected by the presence of a number of serum substances, tested as potential interferences. Attachment of enzymes onto an insoluble support not only provides their reusability but also realizes the extra stabilization rendered by the multipoint covalent attachment or multisubunit binding of the enzymes on solid supports. Use of rationally designed nanoscaffolds for enzyme binding, method development, evaluation, and optimization, overview
additional information
a naturally occuring mutation in intron 3 causes the insertion of an additional exon
additional information
-
a naturally occuring mutation in intron 3 causes the insertion of an additional exon
additional information
NR4A11-250, NR4A11-375, NR4A1251-375, NR4A1251-598, and NR4A1376-598 proteins are obtained from whole-cell lysates of HEK293 cells transfected with the corresponding vectors. Selected Gyk isoform b-targeting small interfering RNA (siRNA)1555 oligonucleotides. Gyk inhibits the effects of NR4A1 on hepatic gluconeogenesis in diabetic mice
additional information
silencing using short hairpin RNA (shRNA) vectors against the GK5 genes shGK5-1 and shGK5-2. shGK5-1 and -2 significantly inhibit GK5 expressions. Morphological examination shows that GK5 knockdown reduces the number of PC9R cells transfected with shGK5 compared to those transfected with negative control shRNA for 24 h and then treated with 0.001 mM gefitinib for 72 h. The CCK8 assays reveals that transfection of PC9R cells with either shGK5-1 or -2 enhances gefitinib-induced apoptosis. GK5 knockdown induces PC9R cell apoptosis and cell cycle arrest in the presence of gefitinib
additional information
-
silencing using short hairpin RNA (shRNA) vectors against the GK5 genes shGK5-1 and shGK5-2. shGK5-1 and -2 significantly inhibit GK5 expressions. Morphological examination shows that GK5 knockdown reduces the number of PC9R cells transfected with shGK5 compared to those transfected with negative control shRNA for 24 h and then treated with 0.001 mM gefitinib for 72 h. The CCK8 assays reveals that transfection of PC9R cells with either shGK5-1 or -2 enhances gefitinib-induced apoptosis. GK5 knockdown induces PC9R cell apoptosis and cell cycle arrest in the presence of gefitinib
additional information
recombinant expression of GST-tagged wild-type and mutant KPN00353s in Klebsiella pneumoniae strain MGH 78578. Protein pull-down assay, GST-tagged GlpK binds to His-tagged H65D or His-tagged H65E mutant proteins more strongly than to His-tagged H65R or to His-tagged wild-type KPN00353 and binds weakly to His-tagged H65Q. The binding affinity between GlpK and the His-tagged H110Q mutant protein is similar to that between GlpK and His-tagged wild-type KPN00353. Quantification of intracellular G3P in recombinant Klebsiella pneumoniae overexpressing wild-type or variant KPN00353 proteins. Interaction analysis with wild-type and mutant variants of regulator KPN00353, KPN00353 H65 mutants (H65D, H65E, H65Q, and H65R) overexpression leads to altered intracellular glycerol-3-phosphate concentration in the cells compared to wild-type KPN00353
additional information
-
recombinant expression of GST-tagged wild-type and mutant KPN00353s in Klebsiella pneumoniae strain MGH 78578. Protein pull-down assay, GST-tagged GlpK binds to His-tagged H65D or His-tagged H65E mutant proteins more strongly than to His-tagged H65R or to His-tagged wild-type KPN00353 and binds weakly to His-tagged H65Q. The binding affinity between GlpK and the His-tagged H110Q mutant protein is similar to that between GlpK and His-tagged wild-type KPN00353. Quantification of intracellular G3P in recombinant Klebsiella pneumoniae overexpressing wild-type or variant KPN00353 proteins. Interaction analysis with wild-type and mutant variants of regulator KPN00353, KPN00353 H65 mutants (H65D, H65E, H65Q, and H65R) overexpression leads to altered intracellular glycerol-3-phosphate concentration in the cells compared to wild-type KPN00353
-
additional information
-
recombinant expression of GST-tagged wild-type and mutant KPN00353s in Klebsiella pneumoniae strain MGH 78578. Protein pull-down assay, GST-tagged GlpK binds to His-tagged H65D or His-tagged H65E mutant proteins more strongly than to His-tagged H65R or to His-tagged wild-type KPN00353 and binds weakly to His-tagged H65Q. The binding affinity between GlpK and the His-tagged H110Q mutant protein is similar to that between GlpK and His-tagged wild-type KPN00353. Quantification of intracellular G3P in recombinant Klebsiella pneumoniae overexpressing wild-type or variant KPN00353 proteins. Interaction analysis with wild-type and mutant variants of regulator KPN00353, KPN00353 H65 mutants (H65D, H65E, H65Q, and H65R) overexpression leads to altered intracellular glycerol-3-phosphate concentration in the cells compared to wild-type KPN00353
-
additional information
-
Aqp7-/- knock out mice. Gene replacement targeting strategy is used to inactivate the Aqp7 gene in R1 mouse ES cells. Targeted ES cell clones are injected into blastocysts of C57BL/6J mice. Glycerol kinase expression (mRNA level) and activity is increased in pancreatic islets of Aqp7-/- mice compared to Aqp+/+ mice.
additional information
generation of Gk2-7/-7 KO mice using the CRISPR/Cas9 system, analysis of sperm mitochondria in testis using a freeze-fracture method with scanning electron microscopy. To generate the Gk2 KO mice, the pX330 plasmid is prepared expressing a chimeric sgRNA together with human codon-optimized Cas9 (hCas9) by ligating oligonucleotides into the BbsI site. The sgRNAs recognize sequences close to the start codon. Gk2-disrupted spermatids show abnormal localization of crescent-like mitochondria, in spite of the initial proper alignment of spherical mitochondria around the flagellum, which causes abnormal mitochondrial sheath formation leading to exposure of the outer dense fibers. Phenotype of Gk2-7/-7 mice, overview
additional information
-
generation of Gk2-7/-7 KO mice using the CRISPR/Cas9 system, analysis of sperm mitochondria in testis using a freeze-fracture method with scanning electron microscopy. To generate the Gk2 KO mice, the pX330 plasmid is prepared expressing a chimeric sgRNA together with human codon-optimized Cas9 (hCas9) by ligating oligonucleotides into the BbsI site. The sgRNAs recognize sequences close to the start codon. Gk2-disrupted spermatids show abnormal localization of crescent-like mitochondria, in spite of the initial proper alignment of spherical mitochondria around the flagellum, which causes abnormal mitochondrial sheath formation leading to exposure of the outer dense fibers. Phenotype of Gk2-7/-7 mice, overview
additional information
frameshift mutations in a hypervariable homopolymeric region of the glpK gene are a specific marker of multidrug resistance in clinical Mycobacterium tuberculosis isolates, and these loss-of-function alleles are also enriched in extensively drug-resistant clones. Reversible high-frequency variation in carbon metabolic pathways can produce phenotypically drug-tolerant clones and have a role in the development of resistance. Construction of a DELTAglpK mutant, that is significantly less sensitive to isoniazid (INH) and rifampin (RIF) than the wild-type or the complemented mutant
additional information
-
frameshift mutations in a hypervariable homopolymeric region of the glpK gene are a specific marker of multidrug resistance in clinical Mycobacterium tuberculosis isolates, and these loss-of-function alleles are also enriched in extensively drug-resistant clones. Reversible high-frequency variation in carbon metabolic pathways can produce phenotypically drug-tolerant clones and have a role in the development of resistance. Construction of a DELTAglpK mutant, that is significantly less sensitive to isoniazid (INH) and rifampin (RIF) than the wild-type or the complemented mutant
additional information
-
frameshift mutations in a hypervariable homopolymeric region of the glpK gene are a specific marker of multidrug resistance in clinical Mycobacterium tuberculosis isolates, and these loss-of-function alleles are also enriched in extensively drug-resistant clones. Reversible high-frequency variation in carbon metabolic pathways can produce phenotypically drug-tolerant clones and have a role in the development of resistance. Construction of a DELTAglpK mutant, that is significantly less sensitive to isoniazid (INH) and rifampin (RIF) than the wild-type or the complemented mutant
-
additional information
overexpression of glycerol kinase under oxidative stress with glycerol supplementation leads to enhancement of lipid production in Synechocystis sp. PCC 6803
