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Literature summary for 3.2.1.114 extracted from

  • Strasser, R.; Schoberer, J.; Jin, C.; Gloessl, J.; Mach, L.; Steinkellner, H.
    Molecular cloning and characterization of Arabidopsis thaliana Golgi alpha-mannosidase II, a key enzyme in the formation of complex N-glycans in plants (2006), Plant J., 45, 789-803.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
GMII, DNA and amino acid sequence determination and analysis, expression of His-tagged functional Golgi alpha-mannosidase II as soluble protein, which is secreted to the culture medium, in Spodoptera frugiperda Sf21 insect cells using the baculovirus transfection system, transient expression of GFP-tagged enzyme in Nicotiana benthamiana plants shows localization of the recombinant enzyme to the Golgi and endoplasmic reticulum Arabidopsis thaliana

Protein Variants

Protein Variants Comment Organism
additional information construction of two Golgi alpha-mannosidase II knockout mutant plants, lines Salk_052443 and Salk_141821, resulting in predominant presence of unprocessed N-glycans but revealing also that alternative routes for N-glycan processing exist in the plants, mutant substrate specificity analysis, overview Arabidopsis thaliana

Inhibitors

Inhibitors Comment Organism Structure
Cu2+ over 97% inhibition of the recombinant GMII at 1 mM Arabidopsis thaliana
additional information no inhibition of recombinant GMII by EDTA Arabidopsis thaliana
swainsonine 50% inhibition of recombinant GMII at 18 nM Arabidopsis thaliana

Localization

Localization Comment Organism GeneOntology No. Textmining
Golgi apparatus Golgi alpha-mannosidase II, the enzyme has a transmembrane domain and a cytoplasmic tail sufficient for localisation of the enzyme to the Golgi complex, retention by lumenal sequences is not involved, retention mechanism Arabidopsis thaliana 5794
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membrane Golgi alpha-mannosidase II is a type II membrane protein Arabidopsis thaliana 16020
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Metals/Ions

Metals/Ions Comment Organism Structure
additional information no effect on recombinant GMII through Ca2+, Co2+, Mn2+, Ni2+, and Zn2+ Arabidopsis thaliana
Zn2+ the recombinant GMII contains zinc in a stoichiometric amount Arabidopsis thaliana

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
Manalpha(1-6)(Manalpha(1-3))Manalpha(1-6)(GlcNAcbeta(1-2)Manalpha(1-3))Manbeta(1-4)GlcNAcbeta(1-4)GlcNAc-1-Asn + H2O Arabidopsis thaliana physiological substrate, conversion by sequential removal of two alpha1,6-linked and alpha1,3-linked mannose residues from the alpha-1,6-branch of the substrate Manalpha(1-6)(GlcNAcbeta(1-2)Manalpha(1-3))Manbeta(1-4)GlcNAcbeta(1-4)GlcNAc-1-Asn + alpha-D-mannose
-
?
additional information Arabidopsis thaliana Golgi alpha-mannosidase II is a key enzyme in the formation of complex N-glycans in plants ?
-
?

Organism

Organism UniProt Comment Textmining
Arabidopsis thaliana Q9LFR0
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-

Posttranslational Modification

Posttranslational Modification Comment Organism
glycoprotein recombinant secreted His-tagged soluble enzyme from culture medium of Spodoptera frugiperda Sf21 insect cells is N-glycosylated ans sensitive to PNGaseF Arabidopsis thaliana

Purification (Commentary)

Purification (Comment) Organism
recombinant secreted His-tagged soluble enzyme from culture medium of Spodoptera frugiperda Sf21 insect cells by nickel affinity chromatography Arabidopsis thaliana

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
6.38
-
purified recombinant GMII expressed in insect cells, substrate 4-nitrophenyl alpha-mannoside Arabidopsis thaliana

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl alpha-D-mannoside + H2O
-
Arabidopsis thaliana 4-nitrophenol + alpha-D-mannose
-
?
Manalpha(1-6)(Manalpha(1-3))Manalpha(1-6)(GlcNAcbeta(1-2)Manalpha(1-3))(Xylbeta(1-4))Manbeta(1-4)GlcNAcbeta(1-4)GlcNAc-1-Asn + H2O
-
Arabidopsis thaliana Manalpha(1-6)(GlcNAcbeta(1-2)Manalpha(1-3))(Xylbeta(1-4))Manbeta(1-4)GlcNAcbeta(1-4)GlcNAc-1-Asn + alpha-D-mannose
-
?
Manalpha(1-6)(Manalpha(1-3))Manalpha(1-6)(GlcNAcbeta(1-2)Manalpha(1-3))Manbeta(1-4)GlcNAcbeta(1-4)GlcNAc-1-Asn + H2O physiological substrate, conversion by sequential removal of two alpha1,6-linked and alpha1,3-linked mannose residues from the alpha-1,6-branch of the substrate Arabidopsis thaliana Manalpha(1-6)(GlcNAcbeta(1-2)Manalpha(1-3))Manbeta(1-4)GlcNAcbeta(1-4)GlcNAc-1-Asn + alpha-D-mannose
-
?
Manalpha(1-6)(Manalpha(1-3))Manalpha(1-6)(GlcNAcbeta(1-2)Manalpha(1-3))Manbeta(1-4)GlcNAcbeta(1-4)GlcNAc-1-Asn + H2O conversion by sequential removal of two alpha1,6-linked and alpha1,3-linked mannose residues from the alpha-1,6-branch of the substrate Arabidopsis thaliana Manalpha(1-6)(GlcNAcbeta(1-2)Manalpha(1-3))Manbeta(1-4)GlcNAcbeta(1-4)GlcNAc-1-Asn + alpha-D-mannose
-
?
Manalpha(1-6)(Manalpha(1-3))Manalpha(1-6)(GlcNAcbeta(1-2)Manalpha(1-3))Manbeta(1-4)GlcNAcbeta(1-4)GlcNAc-pyridylaminoside + H2O
-
Arabidopsis thaliana Manalpha(1-6)(GlcNAcbeta(1-2)Manalpha(1-3))Manbeta(1-4)GlcNAcbeta(1-4)GlcNAc-pyridylaminoside + alpha-D-mannose
-
?
additional information substrate specificity, overview Arabidopsis thaliana ?
-
?
additional information Golgi alpha-mannosidase II is a key enzyme in the formation of complex N-glycans in plants Arabidopsis thaliana ?
-
?

Subunits

Subunits Comment Organism
More the enzyme has a transmembrane domain and a cytoplasmic tail sufficient for localisation of the enzyme to the Golgi complex, retention by lumenal sequences is not involved Arabidopsis thaliana

Synonyms

Synonyms Comment Organism
GmII
-
Arabidopsis thaliana
Golgi alpha-mannosidase II
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Arabidopsis thaliana

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Arabidopsis thaliana

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.8
-
assay at Arabidopsis thaliana