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Literature summary for 3.2.1.113 extracted from

  • Lobsanov, Y.D.; Yoshida, T.; Desmet, T.; Nerinckx, W.; Yip, P.; Claeyssens, M.; Herscovics, A.; Howell, P.L.
    Modulation of activity by Arg407: structure of a fungal alpha-1,2-mannosidase in complex with a substrate analogue (2008), Acta Crystallogr. Sect. D, 64, 227-236.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
DNA of the Penicillium citrinum alpha-1,2 mannosidase gene lacking the signal sequence is cloned downstream of the Aspergillus amylase promoter and of the aspergillopepsin signal sequence. The resulting fungal expression vector pTAPM1 is transfected into Aspergillus oryzae strain MS2 Penicillium citrinum

Crystallization (Commentary)

Crystallization (Comment) Organism
crystals of both the native protein and the protein-inhibitor complex. Superimpositions are performed with LSQMAN. Comparison with human alpha-1,2-mannosidase–thiodisaccharide complex Penicillium citrinum
in complex with the substrate analogue methyl-alpha-D-lyxopyranosyl-(1',2)-alpha-D-mannopyranoside, to 1.95 A resolution. The intact disaccharide spans the ?1/+1 subsites, with the D-lyxoside ring in the -1 subsite in the 1C4 chair conformation.The absence of the C5' hydroxymethyl group on the D-lyxoside moiety results in the side chain of Arg407 adopting two alternative conformations, the minor one interacting with Asp375 and the major one interacting with both the D-lyxoside and the catalytic base Glu409, thus disrupting its function Penicillium citrinum

Inhibitors

Inhibitors Comment Organism Structure
methyl-alpha-D-lyxopyranosyl-(1',2)-alpha-D-mannopyranoside analogue of the minimal disaccharide substrate mannobiose. Cocrystallization and mass spectrometry data suggest that when methyl-alpha-D-lyxopyranosyl-(1',2)-alpha-D-mannopyranoside is incubated with the mannosidase for long periods of time it is cleaved and that methyl-alpha-D-lyxopyranosyl-(1',2)-alpha-D-mannopyranoside behaves as a substrate Penicillium citrinum
additional information comparison with 1-deoxymannojirimycin and kifunensine Penicillium citrinum

Organism

Organism UniProt Comment Textmining
Penicillium citrinum
-
-
-
Penicillium citrinum P31723
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant alpha-1,2-mannosidase is purified from the medium by chromatography on CM-Toyopearl 650M after ammonium sulfate precipitation Penicillium citrinum

Subunits

Subunits Comment Organism
monomer
-
Penicillium citrinum

Synonyms

Synonyms Comment Organism
alpha-1,2-mannosidase
-
Penicillium citrinum
fungal alpha-1,2-mannosidase
-
Penicillium citrinum

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.6
-
methyl-alpha-D-lyxopyranosyl-(1',2)-alpha-D-mannopyranoside
-
Penicillium citrinum