Information on EC 5.4.2.12 - phosphoglycerate mutase (2,3-diphosphoglycerate-independent)

2-[5-amino-2-(4-fluoro-phenyl)-6-oxo-6H-pyrimidin-1-yl]-N-(1-benzyl-2-oxo-2-thiazol-2-yl-ethyl)-acetamide

?

Substrates: -
Products: -

?

3-phospho-D-glycerate

2-phospho-D-glycerate

24 entries

3-Phosphoglycerate

2-Phosphoglycerate

5 entries

additional information

?

-

Staphylococcus saprophyticus

2-phospho-D-glycerate

3-phospho-D-glycerate

32 entries

3-phospho-D-glycerate

2-phospho-D-glycerate

9 entries

additional information

40 entries

-

Ca2+

-

1 mm, 19% of activity with 0.5 mM Mn2+

Cd2+

divalent cation required for activity

Co2+

CoCl2

inhibition by EDTA is reversible by incubation with CoCl2, but not MnCl2, FeSO4, CuSo4, NiCl2, ZnCl2

Mg2+

Mn2+

Zn2+

additional information

1,10-phenanthrolin-5-amine

1,10-phenanthroline

-

3244

1-methyl-2-(methylthio)-1H-benzimidazole-5-carboxamide

54% inhibition at 0.1 mM

1-methyl-2-(methylthio)-N-(5-nitro-1,3-thiazol-2-yl)-1H-benzimidazole-6-carboxamide

47% inhibition at 0.1 mM

1H-benzimidazole-5-carboxylic acid

69% inhibition at 0.1 mM

2,3-Butanedione

Bacillus subtilis

-

3250

2-(4-nitrophenyl)-1H-benzimidazole-7-carboxylic acid

56% inhibition at 0.1 mM

2-cyclohexyl-5-ethylthieno[3,2-d][1,2]thiazol-2-ium

6-chloro-2-(4-nitrophenyl)-1H-benzimidazole-7-carboxylic acid

59% inhibition at 0.1 mM

6-chloro-2-(methylthio)-1H-benzimidazole-5-carboxylic acid

73% inhibition at 0.1 mM

6-chloro-2-(methylthio)-N-(5-nitro-1,3-thiazol-2-yl)-1H-benzimidazole-5-carboxamide

47% inhibition at 0.1 mM

8-hydroxyquinoline 5-sulfonic acid

Cd2+

-

Cu2+

-

diethyl dicarbonate

irreversible inhibition, saturating concentrations of substrate protect

EDTA

EGTA

-

Genzyme-2

Hg2+

-

iodoacetate

-

ipglycermide

-

N-ethylmaleimide

-

NCDS-1

Ni2+

-

p-mercuribenzoate

-

Pb2+

-

phosphate

100 mM, 70% residual activity

Sodium metavanadate

SPB00184

i.e. 2-hydroxyiminoindane-1,3-dione

Tannic acid

TL00638

i.e. 1-methyl-1H-imidazole-4,5-dicarboxylic acid

748834

vanadate

up to 0.1 mM, 90% maximal inhibition. Inhibition may be partially reversed by EDTA

Zn2+

additional information

17 entries

-

2,3-bisphosphoglycerate

2,3-diphosphoglycerate

13 entries

additional information

10 entries

-

0.467

(2R)-3-phosphoglycerate

A0A384KAQ5

at pH 7.5 and 30°C

775115

0.06 - 1.1

2-phospho-D-glycerate

12 entries

0.369 - 1.47

2-phosphoglycerate

4.4

2-[5-Amino-2-(4-fluoro-phenyl)-6-oxo-6H-pyrimidin-1-yl]-N-(1-benzyl-2-oxo-2-thiazol-2-yl-ethyl)-acetamide

pH 7.4, 1 mM Mn2+

0.04 - 5.2

3-phospho-D-glycerate

28 entries

0.324 - 1.1

3-phosphoglycerate

additional information

-

51.3

(2R)-3-phosphoglycerate

A0A384KAQ5

at pH 7.5 and 30°C

775115

2.04 - 199

2-phospho-D-glycerate

1.51 - 3226

3-phospho-D-glycerate

11 entries

additional information

-

7.84 - 36.45

2-phospho-D-glycerate

19.81 - 3226

3-phospho-D-glycerate

0.0383

1,10-phenanthrolin-5-amine

Brugia malayi

Q4VWF8

pH and temperature not specified in the publication

749047

0.0067 - 0.0104

2-cyclohexyl-5-ethylthieno[3,2-d][1,2]thiazol-2-ium

Staphylococcus saprophyticus

0.0609

SPB00184

Leishmania donovani

D7R810

at pH 7.6 and 37°C

748834

0.0373

TL00638

Leishmania donovani

at pH 7.6 and 37°C

0.003

-

0.014

-

0.1

-

0.24

Clostridium perfringens

-

0.49

Micrococcus luteus

-

0.93

Streptomyces coelicolor

-

1

-

100.4

-

1027

-

12

pH 7.0, 50°C

1200

-

1500

methionyl-phosphoglycerate mutase

2000

-

202.5

-

225

256

pH 7.0, 30°C, formation of 3-phospho-D-glycerate

3.3

-

pH 7.0, 50°C

37

23°C

40

pH 7.0, 30°C, formation of 2-phospho-D-glycerate

419

formation of 2-phospho-D-glycerate, pH 7.6, 25°C

500

selenomethionyl-phosphoglycerate mutase

621

-

622.5

-

cytosolic enzyme

71

72

pH 7.0, 30°C, formation of 3-phospho-D-glycerate

83.8

-

plastidic enzyme

93

Brugia malayi

Q4VWF8

pH 7.0, 30°C, formation of 2-phospho-D-glycerate

additional information

5.8 - 6.2

-

formation of 2-phospho-D-glycerate

661689

7

7.1

-

7.3

-

plastidic enzyme, substrate 2-phosphoglycerate

7.4

assay at

7.5

7.5 - 7.8

-

7.5 - 8

-

7.5 - 8.2

for both directions of reaction

7.7 - 8.3

-

7.8

-

8

6

-

about 40% of maximal activity

6 - 8

6 - 8.1

-

6 - 8.7

-

less than 50% of maximal activity above and below

3234

6.8 - 8.8

more than 75% of maximum activity within this range, formation of 2-phospho-D-glycerate

7.7

-

about 70% of maximal activity

30

37

assay at

5.26

calculated

5.5

predicted from amino acid sequence

651058

6

isoelectric focusing

6.3

calculated from amino acid sequence

Wolbachia endosymbiont of Dirofilaria immitis

7.4

F4ZE02

calculated from amino acid sequence

Anaerobium acetethylicum

-

748573

A0A1D3TPC3, A0A1D3TXG7

UniProt

cofactor-independent isoform

-

-

-

-

SwissProt

-

3239, 3249, 3250, 650753

-

Brugia malayi

-

662259, 749047, 772873

Uniprot

Clostridium perfringens

-

-

Cyberlindnera jadinii

-

3239

-

Dirofilaria immitis

Entamoeba histolytica HM1 IMSS

-

UniProt

Methanocaldococcus jannaschii

-

-

-

-

Methanocaldococcus jannaschii DSM 2661

-

SwissProt

-

-

-

-

-

-

-

-

-

UniProt

Oryctolagus cuniculus

-

3239

-

Oryza sativa

rice

3259

-

-

-

-

-

-

3224, 3239, 3249, 677798

-

almond

-

-

-

-

-

Pyrococcus horikoshii

strain OT3

Pyrococcus horikoshii OT-3

-

strain OT3

-

-

3228, 3243

-

-

651716, 734842

Saccharolobus solfataricus DSM 1617

Uniprot

-

734842

Schizosaccharomyces pombe

Uniprot

Saccharomyces cerevisiae

-

3239

-

-

-

potato

-

-

-

-

746646, 747737

Staphylococcus aureus NCTC 8325

UniProt

-

746646, 747737

Staphylococcus saprophyticus

UniProt

-

-

Streptomyces coelicolor

-

-

cofactor-dependent isoform

Uniprot

Wolbachia endosymbiont of Dirofilaria immitis

wheat

3213, 3214, 3215, 3238, 3244, 3252, 3259

-

mung bean

-

-

UniProt

-

3223, 3226, 3234

-

Zymomonas mobilis

-

3241

-

additional information

by sequence alignment found in Arabidopsia thaliana, Oryza sativa, Anthithamnion sp., Escherichia coli, Bacillus subtilis, Bacillus megaterium, Caenorhabditis elegans

-

-

guard cell proteome, overview

-

-

-

-

additional information

-

optimal enzyme production at 37°C

-

-

-

16020

775042

nucleus

-

-

-

Highest Expressing Human Cell Lines

Filter by:

Cell Line Links

Gene Links

evolution

PGAMs that are dependent on (EC 5.4.2.11) or independent of the 2,3-bisphosphoglycerate cofactor are members of two distinct protein families

malfunction

metabolism

Anaerobium acetethylicum

A0A1D3TPC3, A0A1D3TXG7

anaerobic fermentative metabolism of glycerol. Proteome analysis as well as enzyme assays performed in cell-free extracts demonstrate that glycerol is degraded via glyceraldehyde-3-phosphate, which is further metabolized through the lower part of glycolysis leading to formation of mainly ethanol and hydrogen

748573

physiological function

additional information

Acinetobacter baumannii

Staphylococcus aureus subsp. aureus NCTC 8325

100000

Q5GSN3

around 100000 Da, SDS-PAGE, enzyme fused with N-terminal maltose binding protein

694538

183500

Pyrococcus furiosus

-

gel filtration

Methanocaldococcus jannaschii

190600

-

gel filtration

23800

2 * 23800, calculated, 2 * 24000, SDS-PAGE

24000

2 * 23800, calculated, 2 * 24000, SDS-PAGE

26000

Zymomonas mobilis

-

2 * 26000, SDS-PAGE

3241

44100

-

1 * 44100, calculated, 1 * 48000, SDS-PAGE of His-tagged protein

45179

Methanocaldococcus jannaschii

x * 45179, deduced from nucleotide sequence

651716

45200

-

4 * 45200, deduced from nucleotide sequence

45300

Pyrococcus furiosus

-

4 * 45300, deduced from nucleotide sequence

46000

46800

x * 46000-48000, SDS-PAGE, x * 46800, calculated

734842

48000

-

1 * 44100, calculated, 1 * 48000, SDS-PAGE of His-tagged protein

50000 - 60000

Neurospora crassa

-

gel filtration, SDS-PAGE

3237

52000

Zymomonas mobilis

-

gel filtration

3241

54000

-

sedimentation equilibrium centrifugation

56000

57000

58000

-

gel filtration

60000

60557

1 * 60557, deduced from nucleotide sequence

651058

60724

x * 60724, calculated, x * 60000, recombinant protein with His-tag, SDS-PAGE

61000

Priestia megaterium

-

gel filtration, SDS-PAGE

3249

61779

Brugia malayi

-

MALDI-TOF mass spectrometry

747238

63500

-

plastidic enzyme, gel filtration

64000

Zea mays

-

gel filtration, SDS-PAGE

3234

64500

-

cytosolic enzyme, gel filtration

67

gel filtration

?

dimer

monomer

tetramer

additional information

phosphoprotein

protein is composed of two structural and functiional domains, the phosphatase and the transferase. Comparison with the structurally similar domains of in Bacillus stearothermophilus

Bacillus anthracis

Q81X77

678734

enzyme complexed with ipglycermide analogs, Ce-2 Tyr7Phe and Ce-1 C14NHOH, hanging drop vapor diffusion method

772873

crystals of iPGM complexed with 3-phosphoglycerate and Mn2+ at 1.9 A resolution

crystals of wild-type and S62A mutant at 1.4 A resolution

-

652915

methionyl and selenomethionyl phosphoglycerate mutase, vapor drop hanging diffusion, mixing equal volumes of protein solution, i.e. 50 mg/ml protein in 150 mM 3-phosphoglycerate, pH 7.4 and reservoir solution containing 2 M ammonium sulfate, 25 mM zinc acetate, 20 mM cesium chloride, 15 mM 2-mercaptoethanol, 3% polyethylene glycol 200 and 50 mM Tris-HCl, pH 7.4, crystals diffract to 2.5 A resolution

crystals of iPGAM complexed with 3-phosphoglycerate and Co2+, hanging-drop vapor diffusion, crystallization drops contain equal volumes of reservoir and enzyme solution, 5 mg/ml iPGAM, 1.5 mM 3-phosphoglycerate, 50 mM NaCl, 0.01 mM CoCl2 in 20 mM TEA buffer, pH 7.4, crystals diffract to 1.9 A

649202

iPGAM crystallized with 3-phosphoglycerate or 2-phosphoglycerate at high and low cobalt concentrations (4 mM and 0.01 mM), hanging drop vapor diffusion method, using 0.08 M ammonium acetate, 0.04 M trisodium citrate dihydrate pH 6.0, and 24% (w/v) PEG 4000

705169

space group R32, resolution of 2.2 A

Pyrococcus horikoshii

-

hanging drop vapor diffusion method, using 0.2 M NaCl, 0.1 M Bis/Tris pH 6.3 and 25% (w/v) polyethylene glycol 3350

747737

sitting drop vapor diffusion method, using either 0.1 M HEPES pH 7.5, 20% (w/v) polyethylene glycol 10000 or 0.2 M NaCl, 0.1 M bis-Tris pH 6.5, 25% (w/v) polyethylene glycol 3350, at 25°C

-

-

purified recombinant His-tagged enzyme, hanging drop vapour diffusion method, mixing of 0.003 ml of 8 mg/ml protein solution containing 20 mM Tris-acetate-EDTA, pH 7.4, 50 mM NaCl, 001 mM CoCl2, with 0.003 ml of reservoir solution containing 0.05 M ammonium sulfate, 0.1 M Bis-Tris, pH 6.1, and 25% w/v PEG 3350, 18°C, X-ray diffraction structure determination and analysis at 2.3 A resolution

D29N

inactive

R290L

inactive

D12N

no activity

H123N

3% of wild-type activity

H125N

67% of wild-type activity

H128N

20% of wild-type activity

H407N

no activity

H42N

91% of wild-type activity

H42N/H128N

8% of wild-type activity

H445N

18% of wild-type activity

H462N

0.3% of wild-type activity

H66N

2% of wild-type activity

R261L

no activity

S62A

H23A

no enzymic activity. Like wild-type, mutant forms a dimer

D319A

site-directed mutagenesis, substitution of the metal-binding residue Asp319 by Ala results in complete loss of independent PGAM activity

additional information

11

-

24 h, 4°C, 15% loss of activity

5 - 9.5

-

5.5

-

8 h, 4°C, complete inactivation

6 - 10

-

6.8

-

rapid inactivation

50

-

plastidic enzyme: 50 min, 50% loss of activity, cytosolic enzyme: 125 min, 50% loss of activity, no protection by 2-phosphoglycerate, 3-phosphoglycerate, 2,3-bisphosphoglycerate, phosphate

60

70

-

half-life 150 min in presence of Mn2+ and Mg2+. Half-life 30 min in the absence of Mn2+ and Mg2+

85

-

half-life 15 min in presence of Mn2+ and Mg2+

PMSF increases the stability during purification procedure

Neurospora crassa

-

3237

rapid decrease in activity when no stabilizer is added. In presence of NaCl, CoCl2, and imidazole, stable for 1 month at 4°C

-20°C

-

-20°C, 10 mM Tris-HCl buffer, pH 8.0

Aspergillus nidulans

-

3238

-20°C, 50% v/v glycerol, less than 50% loss of activity in 2 months

-

661689

-20°C, 6 months, 10% loss of activity

Zea mays

-

3234

-20°C, potassium phosphate buffer, pH 7.2, 20-25% glycerol, 1 month, no loss of activity

-

0-4°C, ammonium sulfate suspension, pH 7.2

-

3252

-

10 entries

amylose column chromatography

Q5GSN3

694538

His-Select nickel affinity resin column chromatography

A0A384KAQ5

775115

HisTrap column chromatography and Sephadex gel filtration

5 entries

HiTrap nickel chelating column chromatography and HiTrap Q column chromatography

metal affinity Talon column chromatography

705169

Ni-NTA agarose column chromatography

Ni-NTA bead chromatography

Leishmania donovani

-

748833

Ni-NTA column chromatography

Ni-NTA Sepharose column chromatography and Superdex 200 gel filtration

746646

partially purified

651716

partialy purified

7 entries

recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by metal affinity chromatography and gel filtration

recombinant His6-tagged enzyme from Escherichia coli strain C2566/T7 by nicke affinity chromatography, dialysis and anion exchange chromatography

-

recombinant His6-tagged enzyme from Escherichia coli strain C2566/T7 by nickel affinity chromatography, dialysis and anion exchange chromatography

recombinant iPGAM

recombinant iPGM

recombinant phosphoglycerate mutase

recombinant protein

recombinant protein expressed in Escherichia coli

Pyrococcus horikoshii

-

-

4 entries

expressed in Escherichia coli (inactive unsoluble protein) and Kluyveromyces lactis (active enzyme)

Q5GSN3

694538

expressed in Escherichia coli BL21(DE3) cells

7 entries

expressed in Escherichia coli BL21-Codon Plus (DE3) cells

Leishmania donovani

D7R810

748834

expressed in Escherichia coli BLR(DE3) cells

expressed in Escherichia coli C2566/T7 cells

expressed in Escherichia coli M15 (pREP4) cells

746646

expression in Escherichia coli

11 entries

expression in Escherichia coli with C-terminal His-tag

expression in Escherichia coli with His-tag

-

expression in Escherichia coli with His6-tag

expression of C-terminally His-tagged enzyme in Escherichia coli strain C2566/T7

-

expression of C-terminally His6-tagged enzyme in Escherichia coli strain C2566/T7

Dirofilaria immitis

F4ZE01

expression of native and selenomethionyl phosphoglycerate mutase in Escherichia coli

His-tagged enzyme is expressed in Escherichia coli BL21 cells

705169

recombinant expression of the His-tagged enzyme in Escherichia coli strain BL21(DE3)

compared to the wild type plants, at 20°C, the enzyme expression drastically increases in the tcm12 mutant, but at 32°C it is somewhat down-regulated

-

775042

inactivation by EDTA is reversible by incubation with CoCl2, but not MnCl2, FeSO4, CuSo4,NiCl2, ZnCl2

inhibition of enzyme by vanadate may be partially reversed by EDTA

metal ions do not reactivate the enzyme after denaturation with guanidine-HCl

reactivation of EDTA-treated enzyme by Mn2+. Several activation-deactivation cycles can be elicited

Priestia megaterium

-

677798

biofuel production

Anaerobium acetethylicum

A0A1D3TPC3, A0A1D3TXG7

proteome analysis as well as enzyme assays performed in cell-free extracts demonstrates that glycerol is degraded via glyceraldehyde-3-phosphate, which is further metabolized through the lower part of glycolysis leading to formation of mainly ethanol and hydrogen. Fermentation of glycerol to ethanol and hydrogen by this bacterium represents a remarkable option to add value to the biodiesel industries by utilization of surplus glycerol

748573

drug development

TbiPGAM is an attractive molecular target for drug development, the apoenzyme conformation described here provides opportunities for its use in structure-based drug design approaches

medicine