Information on EC 2.4.1.186 - glycogenin glucosyltransferase

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Substrates: recombinant enzyme expressed in E. coli, 71% activity compared to UDP-glucose
Products: -

?

CDP-glucose + p-nitrophenyl-alpha-maltoside

CDP + ?

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Substrates: recombinant enzyme expressed in E. coli
Products: -

?

TDP-glucose + glycogenin

TDP + glucosylated glycogenin

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Substrates: recombinant enzyme expressed in E. coli, 33% activity compared to UDP-glucose
Products: -

?

TDP-glucose + p-nitrophenyl-alpha-maltoside

TDP + ?

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Substrates: recombinant enzyme expressed in E. coli
Products: -

?

UDP-alpha-D-glucose + glycogenin

UDP + alpha-D-glucosylglycogenin

10 entries

UDP-alpha-D-glucose + glycogenin-1

UDP + alpha-D-glucosylglycogenin-1

UDP-alpha-D-glucose + glycogenin-2

UDP + alpha-D-glucosylglycogenin-2

Substrates: self-glucosylation of the glycosyltransferase glycogenin-2
Products: -

?

UDP-galactose + glycogenin

UDP + galactosylated glycogenin

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638247

Substrates: autoglycosylation reaction
Products: -

?

UDP-glucose + glycogenin

UDP + glucosylated glycogenin

44 entries

UDP-glucose + glycogenin

UDP + glucosylglycogenin

UDP-glucose + maltose

UDP + maltotriose

Substrates: trans-glucosylation. 93% of the transferred glucose molecules appears in maltotriose, 6% are attached to glycogenin, and 1% is liberated as free glucose
Products: -

?

UDP-glucose + N-(maltosyl-alpha-1,4-(1-deoxyglucitol))-peptide

UDP + glucosylated N-(maltosyl-alpha-1,4-(1-deoxyglucitol))-peptide

UDP-glucose + n-dodecyl-beta-D-maltoside

UDP + n-dodecyl-beta-D-maltotriose

11 entries

UDP-glucose + n-octyl-alpha-D-maltoside

?

UDP-glucose + n-octyl-beta-D-maltoside

?

UDP-glucose + n-tetradecyl-beta-D-maltoside

?

UDP-xylose + glycogenin

UDP + xylosylated glycogenin

5 entries

UDP-xylose + n-dodecyl-beta-D-maltoside

?

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Substrates: transglucosylation reaction
Products: -

?

additional information

?

-

12 entries

UDP-alpha-D-glucose + glycogenin

UDP + alpha-D-glucosylglycogenin

9 entries

UDP-alpha-D-glucose + glycogenin-1

UDP + alpha-D-glucosylglycogenin-1

Substrates: self-glucosylation of the glycosyltransferase glycogenin-1
Products: -

?

UDP-alpha-D-glucose + glycogenin-2

UDP + alpha-D-glucosylglycogenin-2

Substrates: self-glucosylation of the glycosyltransferase glycogenin-2
Products: -

?

UDP-glucose + glycogenin

UDP + glucosylated glycogenin

14 entries

UDP-glucose + glycogenin

UDP + glucosylglycogenin

additional information

?

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Mg2+

Mn2+

11 entries

ADP

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skeletal muscle enzyme, allosteric inhibition of autoglucosylation

ATP

CDP

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renal enzyme, 90% inhibition at 0.025 mM

CDP-choline

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renal enzyme, 75% inhibition at 0.1 mM

CDP-glucose

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EDTA

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maltose

phosphate

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slight inhibition at 100 mM

TDP-glucose

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UDP

UDP-xylose

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competitive inhibitor to glucosylation of glycogenin by UDP-glucose

additional information

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additional information

Coturnix sp.

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enzyme is phosphorylated in embryonic muscle

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0.1

n-dodecyl-beta-D-maltoside

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3

p-nitrophenyl-alpha-maltoside

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0.002 - 0.00441

UDP-glucose

additional information

5 entries

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0.0053 - 0.0183

UDP-glucose

additional information

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the catalytic effectiveness of glycogenin falls off dramatically as the average glucosyl chain length increases

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additional information

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7

7.4

assay at

7.5

30

5.05

sequence calculation

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Caenorhabditis elegans

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737167

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Coturnix sp.

Quail

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-

hen

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UniProt

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SwissProt

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Hampshire crossbred animals with and without presence of th RN- allele. The total muscle glucose content 0.5 h post-mortem is 77% higher in RN- carriers compared with wild type animals. A greater transcription of glycogenin is found in RN- carriers. The glycogenin m-RNA is more abundant in RN- carriers compared with wild type animals.

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CgGN mRNA expression pattern in adductor muscle during different months, overview. CgGN mRNA expression is closely related to glycogen content and CgGS expression

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Coturnix sp.

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CgGN mRNA expression patterns in gonadal tissue during different months, overview

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the samples for gene expression analyses are taken 0.5 h after slaughter

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-

-

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additional information

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additional information

Highest Expressing Human Cell Lines

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Cell Line Links

Gene Links

evolution

malfunction

10 entries

physiological function

14 entries

additional information

103000

wild-type enzyme in glycosylated and nonglycosylated form, full-length mutant enzymes D162S and D159S, gel filtration

124000

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gel filtration, heterodimeric glycogen synthase complex

488404

200000

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gel filtration, active proteoglycogen

638247

31000

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SDS-PAGE

32000

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kidney enzyme

34000

non-glycosylated truncated protein, gel filtration of a 0.0005 mM solution

690731

37280

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amino acid sequence determination

38000

40000

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recombinant His-tagged unglucosylated glycogenin-1, gel filtration

41000

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recombinant His-tagged autoglucosylated glycogenin-1, gel filtration

47000

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a glycogenin species of average molecular weight 47000 Da is isolated from proteoglycogen isoamylolyzed for 4.5 h, SDS-PAGE

702021

58000

truncation mutant enzymes DELTA270-332/D159S and DELTA270-332/D162S, gel filtration

59000

non-glycosylated truncated protein, gel filtration of a 0.02 mM solution

?

dimer

monomer

tetramer

mutant enzymes D159N, D162N and DELTA270-332/D162N exist as both tetrameric and dimeric species

additional information

17 entries

glycoprotein

19 entries

glycogen synthase-glycogenin enzyme complex, PDB ID 4QLB, GS-N interaction surface, overview

Caenorhabditis elegans

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737167

purified recombinant hGYG1 in complex with Mn2+ and UDP-alpha-glucose, 10 mg/ml hGYG1 with various ligands by sitting drop vapor diffusion at 20°C, X-ray diffraction structure determination and analysis at 2.6 A, molecular replacement

723645

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659695

purified recombinant glycogenin-1 truncation mutant DELTA-270, hanging drop vapor diffusion method, the protein in 12% w/v, PEG monomethyl ester 5000, 0.1 M MES, pH 6.5, and 0.2 M ammonium sulfate, is mixed with 1 M ammonium sulfate and 0.1 M sodium phosphate, pH 6.7, 4°C, crystals are soaked in mother liquor containing 1 mM MnSO4 and 1 mM UDP or UDP-glucose, X-ray diffraction structure determination and analysis

the DELTA270 enzyme crystallizes at 7.5 mg/ml from solutions containing 1 mM UDP-glucose, 1 mM MnCl2, 100 mM sodium acetate, pH 4.54.7, and 1013% (w/v) PEG 4000 in the space group P64, with cell dimensions a = b = 75.0, c = 233.4, gamma = 120° and a dimer in the asymmetric unit. All crystals are grown using hanging drop vapor diffusion methods at 23°C

Y194F

Caenorhabditis elegans

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site-directed mutagenesis, inactive mutant

D163T

D56H/D160T

naturally occuring mutations c.166G.C (p.Asp56His)/c.472Del (p.Asp160Thr fs*5) in GYG1 causing glycogen storage disease (GSD) type XV, phenotype, overview

759550

G135R

determination of a naturally occuring missense mutation that causes reduced expression of glycogenin-1 protein and abolishes the enzyme's activity and function, phenotype includes altered morphology, muscle weakness and wasting, overview

758611

T83A

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site-directed mutagenesis, the mutant shows no incorporation of glucose, no autoglycosylation

T83C

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site-directed mutagenesis, the mutant shows no incorporation of glucose, no autoglycosylation

T83F

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site-directed mutagenesis, the mutant shows no incorporation of glucose, no autoglycosylation

T83M

T83S

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site-directed mutagenesis, the mutant is catalytically active

T83V

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site-directed mutagenesis, the mutant shows no incorporation of glucose, no autoglycosylation

T83Y

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site-directed mutagenesis, the mutant shows no incorporation of glucose, no autoglycosylation

Y195F

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site-directed mutagenesis, glycogenin-1 with the Thr83Met substitution is unable to form the glucose-O-tyrosine linkage at tyrosine 195 unless co-expressed with the catalytically active Tyr195Phe glycogenin-1

DELTA306-664

7fold increase in self-glucosylation

DELTA306-664/Y196F

no self-glucosylation activity

DELTA306-664/Y196F/Y198F

expression results in no accumulation of glycogen

DELTA360-664

11.8fold increase in self-glucosylation

DELTA360-664/Y196F

expression results in reduced glycogen accumulation to 30% of the wild-type enzyme, very low self-glucosylation activity

DELTA360-664/Y196F/Y198F

no self-glucosylation activity

D159N

exists as both tetrameric and dimeric species, compared to wild-type enzyme which exists to more than 95% as dimer, self-glucosylation activities below the limit of detection of the assay. Ability to catalyze the transglucosylation of maltose is reduced by 260fold, hydrolysis of UDP-glucose is reduced 12fold

D159S

stable enzyme, self-glucosylation activities below the limit of detection of the assay. Transglucosylation activity of the mutant enzyme is reduced to undetectable levels, activity for the hydrolysis of UDP-glucose is reduced 14fold

D162N

exists as both tetrameric and dimeric species, compared to wild-type enzyme which exists to more than 95% as dimer, self-glucosylation activities below the limit of detection of the assay, undetectable activity for the transglucosylation of maltose and the hydrolysis of UDP-glucose to free glucose

D162S

stable enzyme, self-glucosylation activities below the limit of detection of the assay. 30fold less active for the trans-glucosylation of maltose and 340fold less active for the hydrolysis of UDP-glucose

DELTA270-332

mutant enzyme is fully active, specific activity for self- or transglucosylation is indistinguishable from the full-length enzyme

DELTA270-332/D159S

inactive mutant enzyme

DELTA270-332/D162N

exists as both tetrameric and dimeric species, compared to wild-type enzyme which exists to more than 95% as dimer

DELTA270-332/D162S

18fold less active for the transglucosylation of maltose and 190fold less active for the hydrolysis of UDP-glucose than wild-type enzyme, activity for the hydrolysis of UDP-glucose is reduced 4fold

T82M

inactive mutant, the mutation is equivalent to T83M according to previous authors amino acid numbering, it causes glycogenosis showing the loss of Thr82 hydrogen bond to Asp162, the residue involved in the activation step of the glucose transfer reaction mechanism. Autoglucosylation, maltoside transglucosylation and UDP-glucose hydrolyzing activities are abolished

T83S

site-directed mutagenesis, the mutant is catalytically active

T83V

site-directed mutagenesis, inactive mutant

Y194F

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exchange of glucose attachment site, no autoglucosylation activity

637622, 638249

Y194X

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mutation at Y194 leads to a protein unable to attach glucose to itself

659125

Y194F

Y194T

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exchange of glucose attachment site, no autoglucosylation activity, mutant glycosylates other substrates but with less activity compared to the wild-type

additional information

9 entries

alkali-stable

concentrated to 8 mg/ml, and 100 microl aliquots are flash frozen in liquid nitrogen and stored at -80°C. Glycogenin stored in this manner is stable for at least 2 years.

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loss of activity on Q-Sepharose during ion exchange chromatography

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-20°C, amylolyzed M-glycogenin preparation, at least 1 month without appreciable loss of activity

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4°C, amylolyzed M-glycogenin preparation, at least 1 week without appreciable loss of activity

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Ni-Sepharose column chromatography and Q Sepharose column chromatography

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702021

nickel-NTA column and Q-Sepharose column

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recombinant GST-tagged glycogen synthase in complex with glycogenin-1 from Sf9 insect cells by glutathione affinity chromatography and gel filtration

737192

recombinant His-tagged wild-type and mutant enzymes from cell-free expression by nickel affinity chromatography

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recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chrmatography

723645

recombinant His6-tagged wild-type and mutant glycogenin-1 from HEK-293 cells by nickel affinity chromatography

recombinant His6-tagged wild-type and mutant glycogenin-2 from HEK-293 cells by nickel affinity chromatography

recombinant proteins

690731

recombinant wild-type gycogenin-1, its truncated mutant DELTA-270, and of glycogenin-1 point mutants from Escherichia coli strain CGSC 4997

separation from glycogen synthase, EC 2.4.1.11, by LiBr

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488404, 637622, 638245

separation from proteoglycogen

wild-type and mutant enzymes

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cell-free expression of His-tagged wild-type and mutant enzymes

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expressed in C2C12 cells

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703952

expressed in Escherichia coli strain Rosetta (DE3)

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702021

expression in COS cells

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expression in Escherichia coli as a GST-tagged protein

expression of glucose-free apo-glycogenin in an Escherichia coli mutant lacking UDP-glucose, enzyme is active towards itself and other substrates

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

723645

expression of mutant Y194F in Escherichia coli

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637622, 638249

expression of wild-type and mutant enzymes in Saccharomyces cerevisiae

expression of wild-type gycogenin-1, its truncated mutant DELTA-270, and of glycogenin-1 point mutants in Escherichia coli strain CGSC 4997

expresssion of recombinant protein in the CGSC Escherichia coli cell line 4997, which lacks UDP-glucose pyrophosphorylase activity

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functional expression of the wild-type enzyme in Escherichia coli

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638249

gene CGI_10023521, DNA and amino acid sequence determination and analysis, the Pacific oyster encodes three isozymes of glycogenin: CgGN-alpha, CgGN-beta, and CgGN-gamma, genetic structure and sequence comparisons, phylogenetic tree. The isolated three CgGN isoforms contain alternative exon regions. PCR-based quantitative expression analysis. Recombinant expression of FLAG-tagged enzymes in HEK-293T cells, recombinant expression of EGFP-tagged enzyme from pEGFP-N1-CgGN in HeLa cells

Magallana gigas

K1R9A7

759384

gene GYG-1, genotyping

758611

gene GYG1, genotyping

O15488

759550

gene GYG1, recombinant expression of functional GST-tagged human muscle glycogen synthase in complex with human glycogenin-1 in Spodoptera frugiperda Sf9 cells using a bicistronic pFastBac-Dual expression vector and baculovirus transfection

737192

gene GYG2, genotyping

isolation of cDNA

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preparation of a truncated already glucosylated His-tag fusion protein in Escherichia coli ER2566, expression of the truncated non-glucosylated His-tag fusion protein in Escherichia coli CGSC 4997

690731

recombinant expression of His6-tagged wild-type and mutant enzymes glycogenin-1 in HEK-293 cells. Coexpression of wild-type glycogenin-2 together with different glycogenin-1 variants using a cell-free system, coexpression of functional glycogenin-1 and glycogenin-2 increases the glucosylation of glycogenin-2