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Literature summary extracted from

  • Numao, S.; Kuntz, D.A.; Withers, S.G.; Rose, D.R.
    Insights into the mechanism of Drosophila melanogaster Golgi alpha-mannosidase II through the structural analysis of covalent reaction intermediates (2003), J. Biol. Chem., 278, 48074-48083.
    View publication on PubMed

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.2.1.24 hanging drop vapour diffusion method, crystallization of wild-type enzyme in complex with 5-fluoro-beta-L-gulosyl fluoride, mutant enzyme D341N in complex with 2-deoxy-2-fluoro-alpgha-D-mannosyl fluoride and mutant enzyme D341N in complex with 5-fluoro-beta-L-gulosyl fluoride Drosophila melanogaster
3.2.1.114 wild-type and D341N mutant GMII, hanging drop vapor diffusion Drosophila melanogaster

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.24 D341N the turnover numberis lower by roughly 200fold compared with that of wild-type enzyme Drosophila melanogaster
3.2.1.114 D341N mutant with removed acid/base catalyst, 2fold lower Km and 200fold kcat values compared with the wild-type enzyme Drosophila melanogaster

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.24 2-deoxy-2-fluoro-alpha-D-mannosyl fluoride for mutant enzyme D341N, no inhibition of wild-type enzyme Drosophila melanogaster
3.2.1.24 5-fluoro-beta-L-gulosyl fluoride reversible, acts as a slow substrate for the D341 mutant enzyme, with deglycosylation as the rate-limiting step. Inactivation is only observed when assayed at low temperatures such that deglycosylation is slow relative to the assay time Drosophila melanogaster
3.2.1.114 2-deoxy-2-fluoro-alpha-D-mannosyl fluoride reversible, D341N mutant GMII Drosophila melanogaster
3.2.1.114 5-fluoro-beta-L-gulosyl fluoride reversible, wild-type and D341N mutant GMII, inhibits only at low assay temperatures, acts as slow substrate at 37°C Drosophila melanogaster
3.2.1.114 swainsonine
-
 Drosophila melanogaster

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.24 additional information
-
 additional information the KM-value for 2-deoxy-2-fluoro-alpha-D-mannosyl fluoride is above 25 mM, wild-type enzyme Drosophila melanogaster
3.2.1.24 0.2
-
 5-fluoro-beta-L-gulosyl fluoride wild-type enzyme Drosophila melanogaster
3.2.1.24 5
-
 2,4-dinitrophenyl alpha-D-mannoside wild-type enzyme Drosophila melanogaster
3.2.1.114 0.05
-
 2,4-dinitrophenyl alpha-D-mannoside pH 5.6, 37°C, D341N mutant GMII Drosophila melanogaster
3.2.1.114 0.2
-
 5-fluoro-beta-L-gulosyl fluoride pH 5.6, 37°C, wild-type GMII Drosophila melanogaster
3.2.1.114 5.5
-
 2,4-dinitrophenyl alpha-D-mannoside pH 5.6, 37°C, wild-type GMII Drosophila melanogaster
3.2.1.114 25
-
 2-deoxy-2-fluoro-alpha-D-mannosyl fluoride pH 5.6, 37°C, above, wild-type GMII Drosophila melanogaster

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.2.1.24 Golgi apparatus
-
 Drosophila melanogaster 5794
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.114 Zn2+ active site coordination to Zn2+ Drosophila melanogaster

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.114 120500
-
 x * 120500, D341N mutant GMII Drosophila melanogaster

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.1.114 additional information Drosophila melanogaster asparagine-linked glycoprotein biosynthesis ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.24 Drosophila melanogaster
-
-
-
3.2.1.114 Drosophila melanogaster Q24451
-
-

Reaction

EC Number Reaction Comment Organism Reaction ID
3.2.1.114 Man5GlcNAc3-[protein] + 2 H2O = Man3GlcNAc3-[protein] + 2 alpha-D-mannopyranose catalytic mechanism Drosophila melanogaster
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.24 2,4-dinitrophenyl alpha-D-mannoside + H2O
-
 Drosophila melanogaster 2,4-dinitrophenol + alpha-D-mannose
-
 ?
3.2.1.24 2-deoxy-2-fluoro-alpha-D-mannosyl fluoride + H2O substrate for wild-type enzyme and very slow substrate of mutant enzyme D341N Drosophila melanogaster ?
-
 ?
3.2.1.24 5-fluoro-beta-L-gulosyl fluoride + H2O acts as a slow substrate for the D341 mutant enzyme, with deglycosylation as the rate-limiting step. Inactivation is only observed when assayed at low temperatures such that deglycosylation is slow relative to the assay time Drosophila melanogaster ?
-
 ?
3.2.1.114 2,4-dinitrophenyl alpha-D-mannoside + H2O
-
 Drosophila melanogaster 2,4-dinitrophenol + alpha-D-mannose
-
 ?
3.2.1.114 2-deoxy-2-fluoro-alpha-D-mannosyl fluoride wild-type and D341N mutant GMII, acts as very slow substrate of the mutant enzyme with a rate-limiting deglycosylation step Drosophila melanogaster ?
-
 ?
3.2.1.114 5-fluoro-beta-L-gulosyl fluoride + H2O acts as slow substrate with a rate-limiting deglycosylation step, wild-type and D341N mutant GMII Drosophila melanogaster ?
-
 ?
3.2.1.114 additional information enzyme/active site structure and substrate binding, catalytic mechanism Drosophila melanogaster ?
-
 ?
3.2.1.114 additional information asparagine-linked glycoprotein biosynthesis Drosophila melanogaster ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.114 ? x * 120500, D341N mutant GMII Drosophila melanogaster

Synonyms

EC Number Synonyms Comment Organism
3.2.1.24 Golgi alpha-mannosidase II
-
 Drosophila melanogaster
3.2.1.114 GmII
-
 Drosophila melanogaster
3.2.1.114 More member of glycosyl hydrolase family 38 Drosophila melanogaster

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.114 37
-
 assay at Drosophila melanogaster

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.24 0.0051
-
 5-fluoro-beta-L-gulosyl fluoride wild-type enzyme Drosophila melanogaster
3.2.1.24 0.048
-
 2,4-dinitrophenyl alpha-D-mannoside mutant enzyme D341N Drosophila melanogaster
3.2.1.24 8.6
-
 2,4-dinitrophenyl alpha-D-mannoside wild-type enzyme Drosophila melanogaster
3.2.1.114 0.0051
-
 5-fluoro-beta-L-gulosyl fluoride pH 5.6, 37°C, wild-type GMII Drosophila melanogaster
3.2.1.114 0.048
-
 2,4-dinitrophenyl alpha-D-mannoside pH 5.6, 37°C, D341N mutant GMII Drosophila melanogaster
3.2.1.114 8.6
-
 2,4-dinitrophenyl alpha-D-mannoside pH 5.6, 37°C, wild-type GMII Drosophila melanogaster

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.114 5.6
-
 assay at Drosophila melanogaster

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.2.1.24 0.6
-
 5-fluoro-beta-L-gulosyl fluoride
-
 Drosophila melanogaster
3.2.1.24 7.5
-
 2-deoxy-2-fluoro-alpha-D-mannosyl fluoride
-
 Drosophila melanogaster
3.2.1.114 0.6
-
 5-fluoro-beta-L-gulosyl fluoride pH 5.6, 37°C, D341N mutant GMII Drosophila melanogaster
3.2.1.114 7.5
-
 2-deoxy-2-fluoro-alpha-D-mannosyl fluoride pH 5.6, 37°C, D341N mutant GMII Drosophila melanogaster