Literature summary for 3.2.1.114 extracted from

Suits, M.D.; Zhu, Y.; Taylor, E.J.; Walton, J.; Zechel, D.L.; Gilbert, H.J.; Davies, G.J.
Structure and kinetic investigation of Streptococcus pyogenes family GH38 alpha-mannosidase (2010), PLoS ONE, 5, e9006.

Search for more literature for 3.2.1.114

Cloned(Commentary)

Cloned (Comment)	Organism
gene M1 GAS SF370, DNA and amino acid sequence determination and analysis, cloning of the coding sequence for SpGH38 residues 1-901, overexpression as 3C protease cleavable N-terminal His6-tagged enzyme in Escherichia coli strain BL21(DE3)	Streptococcus pyogenes M1 GAS

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinent enzyme free or in complex with the inhibitor swainsonine, sitting drop vapour diffusion method, mixing of 12 mg/ml protein in 100 mM Tris, pH 8.5, 1.5 M (NH4)2SO4 and 12% v/v glycerol, with reservoir solution, containing 3% v/v glycerol, 54% v/v Tacsimate, pH 7.0, and 2% v/v PEG 6000, also acting as the cryo-protectant, crystals of the swainsonine complex form are obtained by soaking SpGH38 crystals for ,16 h in mother liquor supplemented with 2 mM swainsonine, X-ray diffraction structure determination and analysis at 1.9 A and 2.6 A resolution, respectively	Streptococcus pyogenes M1 GAS

Crystallization (Comment)

Organism

purified recombinent enzyme free or in complex with the inhibitor swainsonine, sitting drop vapour diffusion method, mixing of 12 mg/ml protein in 100 mM Tris, pH 8.5, 1.5 M (NH4)2SO4 and 12% v/v glycerol, with reservoir solution, containing 3% v/v glycerol, 54% v/v Tacsimate, pH 7.0, and 2% v/v PEG 6000, also acting as the cryo-protectant, crystals of the swainsonine complex form are obtained by soaking SpGH38 crystals for ,16 h in mother liquor supplemented with 2 mM swainsonine, X-ray diffraction structure determination and analysis at 1.9 A and 2.6 A resolution, respectively

Streptococcus pyogenes M1 GAS

Inhibitors

Inhibitors	Comment	Organism	Structure
swainsonine	binding structure, overview	Streptococcus pyogenes M1 GAS

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
3.6	-	4-methylumbelliferyl alpha-D-mannopyranoside	pH 6.8, 37°C, recombinant enzyme	Streptococcus pyogenes M1 GAS
27	-	3-O-alpha-D-mannopyranosyl-alpha-D-mannopyranose	pH 6.8, 37°C, recombinant enzyme	Streptococcus pyogenes M1 GAS

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
intracellular	SpGH38 may be intracellular, absence of any classical signal peptide	Streptococcus pyogenes M1 GAS	5622	-

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Zn2+	involved in catalysis	Streptococcus pyogenes M1 GAS

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
high mannose N-glycan + H2O	Streptococcus pyogenes M1 GAS	-	?	-	?
additional information	Streptococcus pyogenes M1 GAS	higher eukaryotic GH38 alpha-mannosides play a key role in the modification and diversification of hybrid N-glycans, processes with strong cellular links to cancer and autoimmune disease	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Streptococcus pyogenes M1 GAS	Q99YP5	gene M1 GAS SF370	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration	Streptococcus pyogenes M1 GAS

Reaction

Reaction	Comment	Organism	Reaction ID
Man5GlcNAc3-[protein] + 2 H2O = Man3GlcNAc3-[protein] + 2 alpha-D-mannopyranose	reaction mechanism via glycosyl-enzyme intermediate, overview	Streptococcus pyogenes M1 GAS	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
2,4-dinitrophenyl alpha-D-mannopyranoside + H2O	-	Streptococcus pyogenes M1 GAS	2,4-dinitrophenol + alpha-D-mannopyranose	-	?
3-O-alpha-D-mannopyranosyl-alpha-D-mannopyranose + H2O	the enzyme is only active with the alpha-1,3-linked disaccharide	Streptococcus pyogenes M1 GAS	alpha-D-mannopyranose	-	?
4-methylumbelliferyl alpha-D-mannopyranoside + H2O	-	Streptococcus pyogenes M1 GAS	4-methylumbelliferol + alpha-D-mannopyranose	-	?
4-nitrophenyl alpha-D-mannopyranoside + H2O	-	Streptococcus pyogenes M1 GAS	4-nitrophenol + alpha-D-mannopyranose	-	?
high mannose N-glycan + H2O	-	Streptococcus pyogenes M1 GAS	?	-	?
additional information	higher eukaryotic GH38 alpha-mannosides play a key role in the modification and diversification of hybrid N-glycans, processes with strong cellular links to cancer and autoimmune disease	Streptococcus pyogenes M1 GAS	?	-	?
additional information	SpGH38 is an alpha-mannosidase with specificity for alpha-1,3 mannosidic linkages, active site structure, overview	Streptococcus pyogenes M1 GAS	?	-	?

Subunits

Subunits	Comment	Organism
More	structure comparisons, overview	Streptococcus pyogenes M1 GAS

Synonyms

Synonyms	Comment	Organism
alpha-1,3 mannosidase	-	Streptococcus pyogenes M1 GAS
alpha-mannosidase	-	Streptococcus pyogenes M1 GAS
GH38 alpha-mannosidase II	-	Streptococcus pyogenes M1 GAS
GH38 enzyme	-	Streptococcus pyogenes M1 GAS
More	the enzyme belongs to the family GH38 of alpha-mannosidases	Streptococcus pyogenes M1 GAS
SpGH38	-	Streptococcus pyogenes M1 GAS

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Streptococcus pyogenes M1 GAS

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.03	-	4-methylumbelliferyl alpha-D-mannopyranoside	pH 6.8, 37°C, recombinant enzyme	Streptococcus pyogenes M1 GAS
6.4	-	3-O-alpha-D-mannopyranosyl-alpha-D-mannopyranose	pH 6.8, 37°C, recombinant enzyme	Streptococcus pyogenes M1 GAS

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
6.8	-	assay at	Streptococcus pyogenes M1 GAS

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.018	-	swainsonine	-	Streptococcus pyogenes M1 GAS

General Information

General Information	Comment	Organism
physiological function	the enzyme is perhaps functioning in the subsequent degradation of extracellular host glycans following their initial digestion by secreted glycosidases	Streptococcus pyogenes

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.01	-	2,4-dinitrophenyl alpha-D-mannopyranoside	pH 6.8, 37°C, recombinant enzyme	Streptococcus pyogenes
0.24	-	3-O-alpha-D-mannopyranosyl-alpha-D-mannopyranose	pH 6.8, 37°C, recombinant enzyme	Streptococcus pyogenes