Information on EC 2.7.1.2 - glucokinase

HgCl2

human glucokinase regulatory protein

inhibition is reversed by activator RO-28-1675

663393

iodoacetamide

ITP

N-acetyl-alpha-D-glucosamine

N-acetylglucosamine

N-ethylmaleimide

p-chlormercuribenzoate

p-hydroxymercuribenzoate

palmitoyl-CoA

glucokinase is inhibited by endogenous long-chain fatty acyl-CoA in islets from omega3-depleted rats

685989

Pb(NO3)2

Trehalose 6-phosphate

A1Z0M6;

TTP

UTP

XTP

in presence of ATP

additional information

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(2R)-3-cyclopentyl-2-[4-(methylsulfonyl)phenyl]-N-(1,3-thiazol-2-yl)propanamide

activator associates with glucokinase in a glucose-independent fashion. Kinetic assays reveal a lag in enzyme progress curves that is systematically reduced when the enzyme is preincubated with the activator. Activator binding is enthalpically driven. The kcat value of glucokinase is almost fully limited by product release, both in the presence and absence of activator

737327

(S)-3-(4-((3-fluoropyrrolidin-1-yl)sulfonyl)phenoxy)-5-((3-methylbut-2-en-1-yl)oxy)-N-(thiazol-2-yl)benzamide

activation: 1.9fold

(S)-3-(4-((3-fluoropyrrolidin-1-yl)sulfonyl)phenoxy)-N-(5-fluorothiazol-2-yl)-5-((3-methylbut-2-en-1-yl)oxy)benzamide

activation: 2.2fold

(S)-6-(3-cyclopentyl-2-[4-(trifluoromethyl)-1H-imidazol-1-yl]propanamido)nicotinic acid

in the presence of liver-specific GKA, progress curves at 1 mM glucose are similar to those at 5 mM, reflecting activation of GK. With steady-state kinetic methods it is shown that there are at least two kinetically distinct forms of glucokinase that interconvert through a slow conformational change and that this interconversion is affected by glucose concentration and a liver-specific GKA

721535

(Z)-2-(4-(cyclopropylsulfonyl)phenyl)-N-(5-(2-methylpropylidene)-4-oxo-4,5-dihydro thiazol-2-yl)-3-(tetrahydro-2Hpyran-4-yl)propanamide

activation: 1.17fold

(Z)-N-(5-benzylidene-4-oxo-4,5-dihydrothiazol-2-yl)-2-(4-(cyclopropylsulfonyl)phenyl)-3-(tetrahydro-2H-pyran-4-yl)propanamide

activation: 1.24fold

1-[6'-(2-hydroxy-2-methylpropoxy)-4-[(5-methylpyridin-3-yl)oxy]-3,3'-bipyridin-6-yl]-3-methylurea

P52789;

i.e. AM-2394. Compound activates glucokinase with an EC50 of 60 nM, increases the affinity of glucokinase for glucose by approximately 10fold, exhibits moderate clearance and good oral bioavailability in multiple animal models, and lowers glucose excursion following an oral glucose tolerance test in an ob/ob mouse model of diabetes

737330

2-(4-(cyclopropylsulfonyl)phenyl)-3-(tetrahydro-2Hpyran-4-yl)-N-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)propanamide

activation: 2.27fold

2-(4-(cyclopropylsulfonyl)phenyl)-N-(4-(prop-1-en-2-yl)thiazol-2-yl)-3-(tetrahydro-2H-pyran-4-yl)propanamide

activation: 2.4fold

2-(4-(cyclopropylsulfonyl)phenyl)-N-(4-isopropylthiazol-2-yl)-3-(tetrahydro-2H-pyran-4-yl)propanamide

activation: 2.97fold

2-(4-(cyclopropylsulfonyl)phenyl)-N-(4-oxo-4,5-dihydrothiazol-2-yl)-3-(tetrahydro-2H-pyran-4-yl)propanamide

activation: 1.65fold

2-(4-(cyclopropylsulfonyl)phenyl)-N-(5,6-dihydro-4Hcyclopenta[d]thiazol-2-yl)-3-(tetrahydro-2H-pyran-4-yl)propanamide

activation: 3.4fold

2-(4-(cyclopropylsulfonyl)phenyl)-N-(5-iodo-4-isopropylthiazol-2-yl)-3-(tetrahydro-2H-pyran-4-yl)propanamide

activation: 1.34fold

2-(4-(cyclopropylsulfonyl)phenyl)-N-(5-methyl-4,5,6,7-tetrahydrothiazolo[5,4-c] pyridin-2-yl)-3-(tetrahydro-2Hpyran-4-yl)propanamide

activation: 2.26fold

2-(4-(methylsulfonyl)phenyl)-N-(4-phenylthiazol-2-yl)-3-(tetrahydro-2H-pyran-4-yl)-propanamide

activation: 1.3fold

2-amino-4-fluoro-5-(1-methyl-1H-imidazol-2-ylsulfanyl)-N-thiazol-2-yl-benzamide

potent synthetic allosteric activator, acts on the wild-type and the N-terminal deletion mutants DELTA N1-11 and DELTA N1-15, mechanism of activation

2-amino-N-(4-methyl-1,3-thiazol-2-yl)-5-[(4-methyl-4H-1,2,4-triazol-3-yl)sulfanyl]benzamide

737327

2-[4-(methylsulfonyl)phenyl]-3-(tetrahydro-2H-pyran-4-yl)-N-(1,3-thiazol-2-yl)propanamide

activation: 2.45fold

3-((3-methylbut-2-en-1-yl)oxy)-5-(4-(morpholinosulfonyl)phenoxy)-N-(thiazol-2-yl)benzamide

activation: 2.1fold

3-(4-(((2S,6R)-2,6-dimethylmorpholino)sulfonyl)phenoxy)-5-((3-methylbut-2-en-1-yl)oxy)-N-(thiazol-2-yl)benzamide

activation: 1.5fold

3-(4-(((2S,6R)-2,6-dimethylmorpholino)sulfonyl)phenoxy)-N-(5-fluorothiazol-2-yl)-5-((3-methylbut-2-en-1-yl)oxy)benzamide

activation: 1.7fold

3-(4-((4-methoxypiperidin-1-yl)sulfonyl)phenoxy)-5-((3-methylbut-2-en-1-yl)oxy)-N-(thiazol-2-yl)benzamide

activation: 1.7fold

3-(4-(8-oxa-3-azabicyclo[3.2.1]octan-3-ylsulfonyl)phenoxy)-5-((3-methylbut-2-en-1-yl)oxy)-N-(thiazol-2-yl)benzamide

activation: 1.7fold

3-(4-(8-oxa-3-azabicyclo[3.2.1]octan-3-ylsulfonyl)phenoxy)-N-(5-fluorothiazol-2-yl)-5-((3-methylbut-2-en-1-yl)oxy)benzamide

activation: 1.9fold

3-(4-(cyclopropylsulfonyl)phenoxy)-5-((3-methylbut-2-en-1-yl)oxy)-N-(4-oxo-4,5-dihydrothiazol-2-yl)benzamide

activation: 1.5fold

3-(4-(cyclopropylsulfonyl)phenoxy)-5-((3-methylbut-2-en-1-yl)oxy)-N-(5-methylpyrazin-2-yl)benzamide

activation: 1.6fold

3-(4-(cyclopropylsulfonyl)phenoxy)-5-((3-methylbut-2-en-1-yl)oxy)-N-(thiazol-2-yl)benzamide

activation: 2.4fold

3-(4-(cyclopropylsulfonyl)phenoxy)-N-(1,5-dimethyl-1H-pyr-azol-3-yl)-5-((3-methylbut-2-en-1-yl)oxy)benzamide

activation: 1.5fold

3-(4-(cyclopropylsulfonyl)phenoxy)-N-(4-(4-fluorophenyl)thiazol-2-yl)-5-((3-methylbut-2-en-1-yl)oxy)benzamide

activation: 0.9fold

3-(4-(cyclopropylsulfonyl)phenoxy)-N-(5-fluorothiazol-2-yl)-5-((3-methylbut-2-en-1-yl)oxy)benzamide

activation: 1.8fold

6-[[3-hydroxy-5-(propan-2-yloxy)benzoyl]amino]pyridine-3-carboxylic acid

737327

N-(4,5-dimethylthiazol-2-yl)-2-(4-(methylsulfonyl)phenyl)-3-(tetrahydro-2H-pyran-4-yl)propanamide

activation: 1.72fold

N-(4-(4-fluorophenyl)thiazol-2-yl)-2-(4-(methylsulfonyl)phenyl)-3-(tetrahydro-2H-pyran-4-yl)propanamide

activation: 1.25fold

N-(4-(4-methoxyphenyl)thiazol-2-yl)-2-(4-(methylsulfonyl)phenyl)-3-(tetrahydro-2H-pyran-4-yl)propanamide

activation: 1.22fold

N-(4-(tert-butyl)thiazol-2-yl)-3-(4-(cyclopropylsulfonyl)phenoxy)-5-((3-methylbut-2-en-1-yl)oxy)benzamide

activation: 0.7fold

N-(4-cyclopropylthiazol-2-yl)-2-(4-(methylsulfonyl)phenyl)-3-(tetrahydro-2H-pyran-4-yl)propanamide

activation: 1.87fold

N-(4-ethylthiazol-2-yl)-2-(4-(methylsulfonyl)phenyl)-3-(tetrahydro-2H-pyran-4-yl)-propanamide

activation: 2.17fold

N-(4-isobutylthiazol-2-yl)-2-(4-(methylsulfonyl)phenyl)-3-(tetrahydro-2H-pyran-4-yl)-propanamide

activation: 2.05fold

N-(4-isopropylthiazol-2-yl)-2-(4-(methylsulfonyl)phenyl)-3-(tetrahydro-2H-pyran-4-yl)-propanamide

activation: 2.49fold

N-(4-tert-butylthiazol-2-yl)-2-(4-(methylsulfonyl)phenyl)-3-(tetrahydro-2H-pyran-4-yl)-propanamide

activation: 1.29fold

N-(5-bromo-4-isopropylthiazol-2-yl)-2-(4-(cyclopropylsulfonyl)phenyl)-3-(tetrahydro-2H-pyran-4-yl)propanamide

activation: 1.67fold

N-(5-fluorothiazol-2-yl)-3-((3-methylbut-2-en-1-yl)oxy)-5-(4-(morpholinosulfonyl)phenoxy)benzamide

activation: 1.8fold

N-(5-fluorothiazol-2-yl)-3-(4-((4-methoxypiperidin-1-yl)sulfonyl)phenoxy)-5-((3-methylbut-2-en-1-yl)oxy)benzamide

activation: 1.7fold

N-(5-isopropyl-4-methylthiazol-2-yl)-2-(4-(methylsulfonyl)phenyl)-3-(tetrahydro-2H-pyran-4-yl)propanamide

activation: 1.13fold

N-(6-fluorobenzo[d]thiazol-2-yl)-2-(4-(methylsulfonyl)phenyl)-3-(tetrahydro-2H-pyran-4-yl)propanamide

activation: 1.3fold

RO-0274375

synthetic activator, activation of wild-type enzyme and mutants V62A, V62T, and V62L, no activation of mutants V62Q, V62E, V62F, and V62K

RO-028165

synthetic activator, activation of wild-type enzyme and mutants V62A, V62T, and V62L, no activation of mutants V62Q, V62E, V62F, and V62K

RO-0283946

synthetic activator, activation of wild-type enzyme and mutants V62A, V62T, and V62L, no activation of mutants V62Q, V62E, V62F, and V62K

RO-28-0450

racemic, activates the enzyme

663393

RO-28-1675

additional information

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1.9

2-deoxy-D-glucose

30°C, pH 7.5, S151A

2-deoxyglucose

30°C, pH 7.5, wild-type

0.5 - 0.67

6-N-(carboxyethyl)ATP

0.55 - 0.65

6-N-(carboxymethyl)ATP

0.38 - 1

6-N-(succinyl)ATP

0.5 - 1.25

6-N-[N-(6-aminohexhyl)carbamoyl]ATP

0.15

alpha-D-glucose

30°C, pH 7.6

0.00000023 - 12.6

ATP

96 entries

0.56

CTP

30°C, pH 7

4.7 - 240

D-fructose

0.028 - 45

D-glucose

48 entries

0.08 - 4.4

D-mannose

6 entries

0.25

GTP

30°C, pH 7

0.36

ITP

30°C, pH 7

0.19

MgATP2-

30°C, pH 7

641069

0.63

UTP

30°C, pH 7

additional information

9 entries

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0.61 - 1492

ATP

5.9 - 166

beta-D-glucose

16 entries

0.007 - 1492

D-glucose

59 entries

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0.01 - 11

D-glucose

6 entries

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0.0041

2-deoxy-2-([[(1,1-dioxido-1-benzothiophen-2-yl)methoxy]carbonyl]amino)-beta-D-glucopyranose

His-tagged recombinant protein, pH 7.2, 22°C

0.0013

2-deoxy-2-[[3-(4-hydroxyphenyl)propanoyl]amino]-alpha-D-glucopyranose

His-tagged recombinant protein, pH 7.2, 22°C

0.00071

2-[[(benzyloxy)carbonyl]amino]-2-deoxy-beta-D-glucopyranose

His-tagged recombinant protein, pH 7.2, 22°C

0.0000128 - 0.000113

GK regulatory protein

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0.0001

fetal glucokinase

0.0003

adult glucokinase

0.0018

strain KT2440, grown on citrate

680495

0.0164

strain KT440 in presence of glucose and toluene

680502

0.0211

strain KT2440, grown on glucose

680495

0.0273

strain KT440 in presence of glucose

0.26

hepatocytes

0.34

S151G, D-mannose as substrate

0.51

Dictyostelium discoideum

641053

0.57

rat hepatocytes treated with adenovirus containing the entire coding sequence of rat liver glucokinase

641071

0.6

S151G, D-glucose as substrate

0.786

substrate: D-fructose

0.941

subatrste: D-mannose

Streptomyces peucetius subsp. caesius

1.09

purified enzyme, Vmax

1.4

N166R-S151A, D-mannose as substrate

1.67

Streptomyces coelicolor

P0A4E1;

purified enzyme, Vmax

1.95

substrate: D-glucose

N166R-S151A, D-glucose as substrate

2.16

S151C, D-mannose as substrate

3.18

S151G, D-fructose as substrate

3.32

S151A, D-mannose as substrate

4.1

S151A, 2-deoxy-D-glucose as substrate

S151C, D-glucose as substrate

6.6

S151A, D-glucose as substrate

9.12

S151C, D-fructose as substrate

14.5

N166R-S151A, D-fructose as substrate

23.8

S151A, D-fructose as substrate

47.2

recombinant protein

60.8

N166R, D-mannose as substrate

wild-type, D-mannose and 2-deoxy-D-glucose as substrate

74.9

wild-type and N166R, D-glucose as substrate

122

wild-type, D-fructose as substrate

158

166

purified recombinant enzyme

198

240

304

310

N166R, D-fructose as substrate

334

370

purified recombinant enzyme, Vmax

additional information

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7.1

7.2

assay at

7.3

7.4

assay at

7.5

7.5 - 9.5

7.6

7.8

8.2

2 optima: a major peak at pH 7.0 and a minor peak at pH 8.2

8.5 - 9

641058, 641060

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6 - 9

Dictyostelium discoideum

pH 6.0: about 55% of activity maximum, pH 9.0: about 75% of activity maximum

641053

6 - 10.5

pH 6.0: about 30% of activity maximum, pH 10.5: about 80% of activity maximum

6.8 - 7.7

50% activity at pH 6.8 and pH 7.7

7 - 8.5

additional information

development of an assay method stable at pH 6.8-8.5 and 75-90°C, overview

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Dictyostelium discoideum

assay at

20 - 25

assay at

assay at

Streptomyces coelicolor

P0A4E1;

Streptomyces peucetius subsp. caesius

Caldanaerobacter subterraneus subsp. tengcongensis

Q8RDE9;

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38 - 98

below 10% of maximal activity at 38°C, about 90% of maximal activity at 98°C

additional information

development of an assay method stable at pH 6.8-8.5 and 75-90°C, overview

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P17712;

O31392;

glk is a nonessential gene that is constitutively expressed during logarithmic growth and has slightly reduced expression in the stationary phase

Q4Q1I9;

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Escherichia coli K-12

661139

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Escherichia coli O157:H7;

30 entries

Homo sapiens;

Lactococcus lactis subsp. lactis (strain IL1403);

6edi, download, 3D-view

SCOPe (6edi)

CATH (6edi)

A4HPA0

Leishmania braziliensis;

3vpz, download, 3D-view

SCOPe (3vpz)

CATH (3vpz)

H7CHS4

Pseudoalteromonas sp. AS-131;

1ua4, download, 3D-view

SCOPe (1ua4)

CATH (1ua4)

Q9V2Z6

Pyrococcus furiosus (strain ATCC 43587 / DSM 3638 / JCM 8422 / Vc1);

4lc9, download, 3D-view

SCOPe (4lc9)

CATH (4lc9)

Q07071

Rattus norvegicus;

Streptomyces griseus subsp. griseus (strain JCM 4626 / NBRC 13350);

Trypanosoma cruzi (strain CL Brener);

3w0l, download, 3D-view

SCOPe (3w0l)

CATH (3w0l)

Q91753

Xenopus laevis;

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24000

Streptococcus mutans

alpha2, 2 * 24000, SDS-PAGE

641054

33000

alpha2, 2 * 33000, SDS-PAGE

641056

33500

2 * 33500, the recombinant enzyme probably is a dimer, SDS-PAGE

34500

2 * 34500, SDS-PAGE

35000

35200

recombinant enzyme, about, mass spectrometry, 2 peaks

36000

2 * 36000, alpha2, recombinant enzyme, SDS-PAGE

41000

gel filtration

43000

x * 43000, the enzyme may occur as a monomer or dimer, dependent on the protein concentration, SDS-PAGE

46000

47000

x * 47000, SDS-PAGE

49000

Q4Q1I9;

monomer, gel filtration

52000

x * 52000, SDS-PAGE

65000

gel filtration

67000

gel filtration

67500

density gradient centrifugation

70000

recombinant enzyme, oxidative conditions, PAGE, the wild-type and mutant enzymes occur as both monomer and homodimer

70400

recombinant enzyme, about, mass spectrometry, 2 peaks

75000

recombinant enzyme, gel filtration

86000

87000

HPLC gel filtration

additional information

recombinant enzyme, native PAGE

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dimer

homodimer

monomer

tetramer

additional information

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glycoprotein

Mus musculus

glucokinase is modified by O-GlcNAcylation. siRNA-mediated O-linked N-acetylglucosamine transferase knock-down not only decreases O-GlcNAc content but also glucokinase protein level

737623

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purified recombinant selenomethionine-labeled enzyme free or in complex with D-glucose, hanging drop vapour diffusion method, 0.002 ml enzyme solution containing 6.8 mg/ml protein mixed with 0.004 ml reservoir solution containing 0.1 M Tris-HCl, pH 8.5, 0.2 M MgCl2, and 1.7 M ammonium sulfate for the apo-enzyme or 18.5-20% PEG 6000 with 2 mM D-glucose and 2-3 mM ADP for the glucose-bound enzyme, X-ray diffraction structure determination and analysis at 2.3-2.2 A resolution

Escherichia coli

P0A6V9;

662022

homology modeling of structure reveals high structural homology with human glucokinase

Felis catus

A3EYB3;

737954

first structures of a glucokinase-glucose complex without activator, of glucokinase-glucose-AMP-PNP and of glucokinase-glucose-AMP-PNP with a bound activator are reported. All structures are extremely similar, thus demonstrating that binding of GK activators does not result in conformational changes of the active protein but in stabilization of the active form of glucokinase

721163

molecular docking of glucose and synthetic ligands (allosteric activators). Predicted position of glucose coincides with its position in the crystallographic complex. Glucose interacts with the residues of large (Ile225, Gly229, Cys230, Asn231, Glu256, Gln287, Glu290) and small (Ser151, Phe152, Pro153, Thr168, Lys169) domains and with the residues of connecting region Asn204, Asp205. Hydrogen bonds with Asp205, Glu256 and Glu290 stabilize this complex. The allosteric site formed by the small and large domains and two loops connecting them is located approximately 20 A away from the glucose binding site. There are two stable positions of glucokinase activators and approximately five sites of ligand binding with less favorable interaction energy. Predicted position of glucose coincides with its position in the crystallographic complex

738083

purified recombinant hepatic wild-type and deletion mutant enzymes in complex with either D-glucose or the synthetic activator 2-amino-4-fluoro-5-(1-methyl-1H-imidazol-2-ylsulfanyl)-N-thiazol-2-yl-benzamide, hanging drop vapour diffusion method, 10 mg/ml protein in 20 mM Tris-HCl, pH 7.5, 50 mM NaCl, 5 mM Tris(2-carboxyethyl)phosphine hydrochloride, and 20 mM D-glucose, or 0.3 mM compound A, 0.0015-0.003 ml of the solution is mixed with an equal volume of precipitant solution containing 28-30% PEG 1500, 0.1 M HEPES-NaOH, pH 6.0, equilibration against 1 ml precipitant solution, 1 week, crystallization of the free hepatic enzyme by using precipitant solution containing 50 mM NaCl, 1.5-1.6 M ammonium sulfate, and 0.1 M bicine-NaOH, pH 8.7, 1 week, X-ray diffraction structure determination and analysis at 2.3 and 3.4 A resolution, respectively, molecular replacement

homology modeling of structure. Staphylococcus aureus glucokinase exhibits very close homology with Enterococcus faecalis and Clostridium difficle while with other bacteria it show a high degree of variations both in domain and nondomain regions. Glucose interacts with residues V77, N114, S161 and G160 forming a total of eight hydrogen bonds. Glucose docking score is -12.3697 kcal/mol

Staphylococcus aureus

H6UH58;

738362

crystal structures of apo-SgGlkA, SgGlkA in complex with glucose, and SgGlkA in complex with glucose and adenylyl imidodiphosphate (AMPPNP) are reported. SgGlkA is divided into a small alpha/beta domain and a large alpha + beta domain, and it forms a dimer-of-dimer tetrameric configuration

Streptomyces griseus

722553

crystallized using the sitting-drop vapour-diffusion method. A crystal of SgGlkA in complex with glucose is obtained and diffracted X-rays to 1.84 A resolution

Streptomyces griseus

721213

crystal structures in complexes with inhibitors 2-[[(benzyloxy)carbonyl]amino]-2-deoxy-beta-D-glucopyranose, 2-deoxy-2-[[3-(4-hydroxyphenyl)propanoyl]amino]-alpha-D-glucopyranose, 2-deoxy-2-([[(1,1-dioxido-1-benzothiophen-2-yl)methoxy]carbonyl]amino)-beta-D-glucopyranose. The analogues all share a common glucose moietythat preserves key enzyme-substrate hydrogen bonding interactions with the monosaccharide hydroxyl groups from C1, C3, C4,and C6. The C2 hydroxyl is replaced by a NH group and also par-ticipates in the key hydrogen bonding

crystals of TcGlcK in complex with D-glucose and ADP are obtained by the hanging-drop, vapor-diffusion method, using PEG3350 as precipitant agent and diammonium hydrogen citrate as additive. A complete native dataset is collected to 2.1 A maximum resolution

688372

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48 h, with glucose and DTT stable, unstable without

stabilization by D-glucose and glycerol is limited since thermal glucokinase denaturation above 40 °C is irreversible even in their presence

1 min, complete loss of activity

no loss of activity in 30 min

about 25% loss of activity in 30 min, presence of 0.01 M glucose + 0.2 M NaCl, about 90% loss of activity, presence of 0.01 M glucose absence of NaCl

quite stable for 120 min

100

additional information

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D-glucose stabilizes the enzyme through a specific, ligand induced intramolecular transition from an open to a closed conformation

DTT stabilizes at 70°C

glucose increases thermal stability

glucose or DTT protects against thermal inactivation at 25°C

glycerol increases the stability

rapid loss of activity in absence of DTT

Streptococcus mutans

641055

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Glycerol

glycerol causes large functional changes and increases stability as a result of generalized restructuring of surface water of glucokinase by an indirect mechanism and these changes can occur with minimal perturbation of the protein folding structure and with preservation of the unique cooperative kinetics of the enzyme whereas D-glucose stabilizes the enzyme through a specific, ligand induced intramolecular transition from an open to a closed conformation

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-20°C, 1 mM DTT, 20-30% glycerol, indefinitely stable

Streptococcus mutans

641055

-20°C, purified recombinant enzyme, 50 mM Tris-HCl, pH 7.5, stable

-20°C, stable for 4 weeks

room temperature, 1 mM DTT, 5 mM MgCl2, stable for 1 week , 4°C, suspended precipitate in a 60% saturated solution of ammonium sulfate, stable for at least 3 months

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8 entries

glutathione S-transferase (GST)-GlkB fusion proteins, wild-type and mutant

Q53Y25;

688144

His-tagged recombinant TcGlcK is expressed in the Escherichia coli BL21 strain and purified to homogeneity by nickel affinity chromatography

688372

large scale

recombinant

recombinant enzyme

recombinant enzyme from Escherichia coli strain BL21(DE3) by heat treatment and Ni2+ chelating affinity chromatography

recombinant FLAG-tagged hepatic wild-type and mutant enzymes from Escherichia coli strain DH5 alpha by ion exchange and glucosamine affinity chromatography, and gel filtration

recombinant His-tagged enzyme from strain BL21(DE3) by nickel affinity chromatography

Escherichia coli

P0A6V9;

662022

recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain RB791 by nickel affinity chromatography

recombinant N-terminally His6-tagged hepatic enzyme from Escherichia coli by nickel affinity chromatography to over 90% purity

recombinant N-terminally His6-tagged pancreatic enzyme from Escherichia coli by nickel affinity chromatography to over 90% purity

recombinant soluble His-tagged enzyme from Escherichia coli strain BL21(DE3) by ultracentrifugation, nickel affinity chromatography, and gel filtration to homogeneity

recombinant wild-type and mutant enzymes from Escherichia coli to homogeneity

660810

recombinant wild-type and mutant V62M and E300K enzymes as GST-fusion proteins by glutathione affinity chromatography to homogeneity

using Ni-NTA chromatography

using Ni-NTA chromatography followed by ion-exchange chromatography on a HiTrap Q FF column and gel filtration on a Superdex 200 column

721163

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expressed in Escherichia coli as a GST-fusion protein

721533, 721735

expressed in Escherichia coli as a GST-tagged fusion protein

722237

expressed in Escherichia coli as a His-tagged fusion protein

expression in Escherichia coli

4 entries

expression in Saccharomyces cerevisiae

Botrytis cinerea

A1Z0M6;

675854

expression of genes nanK, yajF, and ycfX in glk-deficient mutant strain BM5340(DE3)

Escherichia coli K-12

661139

expression of hepatic wild-type and mutant enzymes in Escherichia coli strain DH5 alpha as FLAG-tagged proteins

expression of the N-terminally His6-tagged hepatic enzyme in Escherichia coli

expression of the N-terminally His6-tagged pancreatic enzyme in Escherichia coli

expression of wild-type and mutant enzymes as GST-fusion proteins

expression of wild-type and mutant enzymes in Escherichia coli BL21(DE3)

660810

gene glcK, phylogenetic analysis and tree, functional complementation of enzyme-deficient Escherichia coli strain UE26, overexpression of His-tagged wild-type and mutant enzymes in Escherichia coli strain RB791 as soluble proteins

gene glk is organized in gene clusters, transcriptomic analysis of KT2440 in response to glucose, overview