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Literature summary for 3.2.1.180 extracted from

  • Maruyama, Y.; Nakamichi, Y.; Itoh, T.; Mikami, B.; Hashimoto, W.; Murata, K.
    Substrate spcificity of streptococcal unsaturated glucuronyl hydrolases for sulfated glycosaminoglycan (2009), J. Biol. Chem., 284, 18059-18069.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.2.1.180

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Streptococcus agalactiae
expression in Escherichia coli Streptococcus pneumoniae
expression in Escherichia coli Streptococcus pyogenes

Crystallization (Commentary)

Crystallization (Comment) Organism
hanging-drop vapor-diffusion method, crystal structure at 1.75 A resolution Streptococcus agalactiae

Protein Variants

Protein Variants Comment Organism
D115N no hydrolysis of beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S Streptococcus agalactiae
D175N no hydrolysis of beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S Streptococcus agalactiae
R236A effectively degrades beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S. Almost inactive on beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc4S, although the substrate is degraded by the wild-type enzyme Streptococcus agalactiae
R236H effectively degrades beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S. Almost inactive on beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc4S, although the substrate is degraded by the wild-type enzyme Streptococcus agalactiae

Inhibitors

Inhibitors Comment Organism Structure
2-mercaptoethanol 1 mM, 20% inhibition Streptococcus pyogenes
AlCl3 1 mM, 65% inhibition Streptococcus agalactiae
CoCl2 1 mM, 55% inhibition Streptococcus agalactiae
D-galactose 5 mM, 21% inhibition Streptococcus pyogenes
D-galacturonic acid 5 mM, complete inhibition Streptococcus agalactiae
D-galacturonic acid 5 mM, 99% inhibition Streptococcus pneumoniae
D-GlcA 5 mM, 97% inhibition Streptococcus pneumoniae
D-glucose 5 mM, 40% inhibition Streptococcus pyogenes
D-glucuronic acid 5 mM, 98% inhibition Streptococcus agalactiae
D-glucuronic acid 5 mM, 98% inhibition Streptococcus pyogenes
D-mannose 5 mM, 22% inhibition Streptococcus pyogenes
D-xylose 5 mM, 33% inhibition Streptococcus pyogenes
KCl 1 mM, 22% inhibition Streptococcus agalactiae
L-fucose 5 mM, 50% inhibition Streptococcus pyogenes
L-rhamnose 5 mM, 35% inhibition Streptococcus pyogenes
LiCl 1 mM, 24% inhibition Streptococcus agalactiae
MnCl2 1 mM, 27% inhibition Streptococcus agalactiae
N-acetylglucosamine 5 mM, 17% inhibition Streptococcus agalactiae
NaCl 1 mM, 22% inhibition Streptococcus agalactiae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.18
-
 beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S pH 7.5, 30°C. wild-type enzyme Streptococcus pneumoniae
0.39
-
 beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S pH 7.5, 30°C. wild-type enzyme Streptococcus pyogenes
0.54
-
 beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S pH 7.5, 30°C. wild-type enzyme Streptococcus agalactiae

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
43000
-
 1 * 43000, SDS-PAGE Streptococcus agalactiae
43000
-
 1 * 43000, SDS-PAGE Streptococcus pneumoniae
43000
-
 1 * 43000, SDS-PAGE Streptococcus pyogenes
46079
-
 1 * 46079, calculated from sequence Streptococcus pneumoniae
46100
-
 1 * 46100, calculated from sequence Streptococcus pyogenes
46587
-
 1 * 46587, calculated from sequence Streptococcus agalactiae

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Streptococcus agalactiae the streptococcal unsaturated glucuronyl hydrolases are capable of degrading unsaturated disaccharides derived from mammalian sulfated glycosaminoglycans. This substrate specificity probably enables Streptococci to readily invade mammalian cells through the degradation of extracellular sulfated glycosaminoglycans ?
-
 ?
additional information Streptococcus pneumoniae the streptococcal unsaturated glucuronyl hydrolases are capable of degrading unsaturated disaccharides derived from mammalian sulfated glycosaminoglycans. This substrate specificity probably enables Streptococci to readily invade mammalian cells through the degradation of extracellular sulfated glycosaminoglycans ?
-
 ?
additional information Streptococcus pyogenes the streptococcal unsaturated glucuronyl hydrolases are capable of degrading unsaturated disaccharides derived from mammalian sulfated glycosaminoglycans. This substrate specificity probably enables Streptococci to readily invade mammalian cells through the degradation of extracellular sulfated glycosaminoglycans ?
-
 ?
additional information Streptococcus pyogenes M1 GAS SF370 the streptococcal unsaturated glucuronyl hydrolases are capable of degrading unsaturated disaccharides derived from mammalian sulfated glycosaminoglycans. This substrate specificity probably enables Streptococci to readily invade mammalian cells through the degradation of extracellular sulfated glycosaminoglycans ?
-
 ?
additional information Streptococcus agalactiae NEM316 the streptococcal unsaturated glucuronyl hydrolases are capable of degrading unsaturated disaccharides derived from mammalian sulfated glycosaminoglycans. This substrate specificity probably enables Streptococci to readily invade mammalian cells through the degradation of extracellular sulfated glycosaminoglycans ?
-
 ?

Organism

Organism UniProt Comment Textmining
Streptococcus agalactiae Q8E372
-
-
Streptococcus agalactiae NEM316 Q8E372
-
-
Streptococcus pneumoniae Q8DR77
-
-
Streptococcus pyogenes Q9A0T3
-
-
Streptococcus pyogenes M1 GAS SF370 Q9A0T3
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Streptococcus agalactiae
-
 Streptococcus pneumoniae
-
 Streptococcus pyogenes

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc + H2O 10% of the activity with beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S Streptococcus pyogenes 4-deoxy-L-threo-5-hexosulose-uronate + beta-D-N-acetylgalactosamine
-
 ?
beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc + H2O 16% of the activity with beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S Streptococcus pneumoniae 4-deoxy-L-threo-5-hexosulose-uronate + beta-D-N-acetylgalactosamine
-
 ?
beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc + H2O 3.4% of the activity with beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S Streptococcus agalactiae 4-deoxy-L-threo-5-hexosulose-uronate + beta-D-N-acetylgalactosamine
-
 ?
beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc + H2O 10% of the activity with beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S Streptococcus pyogenes M1 GAS SF370 4-deoxy-L-threo-5-hexosulose-uronate + beta-D-N-acetylgalactosamine
-
 ?
beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc + H2O 3.4% of the activity with beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S Streptococcus agalactiae NEM316 4-deoxy-L-threo-5-hexosulose-uronate + beta-D-N-acetylgalactosamine
-
 ?
beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S + H2O
-
 Streptococcus agalactiae 4-deoxy-L-threo-5-hexosulose-uronate + beta-D-N-acetylgalactosamine-6-O-sulfate
-
 ?
beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S + H2O
-
 Streptococcus pneumoniae 4-deoxy-L-threo-5-hexosulose-uronate + beta-D-N-acetylgalactosamine-6-O-sulfate
-
 ?
beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S + H2O
-
 Streptococcus pyogenes 4-deoxy-L-threo-5-hexosulose-uronate + beta-D-N-acetylgalactosamine-6-O-sulfate
-
 ?
beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S + H2O
-
 Streptococcus pyogenes M1 GAS SF370 4-deoxy-L-threo-5-hexosulose-uronate + beta-D-N-acetylgalactosamine-6-O-sulfate
-
 ?
beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S + H2O
-
 Streptococcus agalactiae NEM316 4-deoxy-L-threo-5-hexosulose-uronate + beta-D-N-acetylgalactosamine-6-O-sulfate
-
 ?
beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GlcNAc + H2O 1.1% of the activity with beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S Streptococcus pyogenes 4-deoxy-L-threo-5-hexosulose-uronate + beta-D-N-acetylglucosamine
-
 ?
beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GlcNAc + H2O 2% of the activity with beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S Streptococcus pneumoniae 4-deoxy-L-threo-5-hexosulose-uronate + beta-D-N-acetylglucosamine
-
 ?
beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GlcNAc + H2O 1.1% of the activity with beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S Streptococcus pyogenes M1 GAS SF370 4-deoxy-L-threo-5-hexosulose-uronate + beta-D-N-acetylglucosamine
-
 ?
additional information the streptococcal unsaturated glucuronyl hydrolases are capable of degrading unsaturated disaccharides derived from mammalian sulfated glycosaminoglycans. This substrate specificity probably enables Streptococci to readily invade mammalian cells through the degradation of extracellular sulfated glycosaminoglycans Streptococcus agalactiae ?
-
 ?
additional information the streptococcal unsaturated glucuronyl hydrolases are capable of degrading unsaturated disaccharides derived from mammalian sulfated glycosaminoglycans. This substrate specificity probably enables Streptococci to readily invade mammalian cells through the degradation of extracellular sulfated glycosaminoglycans Streptococcus pneumoniae ?
-
 ?
additional information the streptococcal unsaturated glucuronyl hydrolases are capable of degrading unsaturated disaccharides derived from mammalian sulfated glycosaminoglycans. This substrate specificity probably enables Streptococci to readily invade mammalian cells through the degradation of extracellular sulfated glycosaminoglycans Streptococcus pyogenes ?
-
 ?
additional information shows almost no activity with the unsaturated gellan tetrasaccharide (beta-D-4-deoxy-DELTA4,5-GlcAp-(1->4)-beta-D-Glcp-(1->4)-alpha-L-Rhap-(1->3)-beta-D-Glcp). Streptococcal enzymes prefer sulfated substrates Streptococcus agalactiae ?
-
 ?
additional information shows almost no activity with the unsaturated gellan tetrasaccharide (beta-D-4-deoxy-DELTA4,5-GlcAp-(1->4)-beta-D-Glcp-(1->4)-alpha-L-Rhap-(1->3)-beta-D-Glcp). Streptococcal enzymes prefer sulfated substrates Streptococcus pneumoniae ?
-
 ?
additional information shows almost no activity with the unsaturated gellan tetrasaccharide (beta-D-4-deoxy-DELTA4,5-GlcAp-(1->4)-beta-D-Glcp-(1->4)-alpha-L-Rhap-(1->3)-beta-D-Glcp). Streptococcal enzymes prefer sulfated substrates Streptococcus pyogenes ?
-
 ?
additional information the streptococcal unsaturated glucuronyl hydrolases are capable of degrading unsaturated disaccharides derived from mammalian sulfated glycosaminoglycans. This substrate specificity probably enables Streptococci to readily invade mammalian cells through the degradation of extracellular sulfated glycosaminoglycans Streptococcus pyogenes M1 GAS SF370 ?
-
 ?
additional information shows almost no activity with the unsaturated gellan tetrasaccharide (beta-D-4-deoxy-DELTA4,5-GlcAp-(1->4)-beta-D-Glcp-(1->4)-alpha-L-Rhap-(1->3)-beta-D-Glcp). Streptococcal enzymes prefer sulfated substrates Streptococcus pyogenes M1 GAS SF370 ?
-
 ?
additional information the streptococcal unsaturated glucuronyl hydrolases are capable of degrading unsaturated disaccharides derived from mammalian sulfated glycosaminoglycans. This substrate specificity probably enables Streptococci to readily invade mammalian cells through the degradation of extracellular sulfated glycosaminoglycans Streptococcus agalactiae NEM316 ?
-
 ?
additional information shows almost no activity with the unsaturated gellan tetrasaccharide (beta-D-4-deoxy-DELTA4,5-GlcAp-(1->4)-beta-D-Glcp-(1->4)-alpha-L-Rhap-(1->3)-beta-D-Glcp). Streptococcal enzymes prefer sulfated substrates Streptococcus agalactiae NEM316 ?
-
 ?

Subunits

Subunits Comment Organism
monomer 1 * 43000, SDS-PAGE Streptococcus agalactiae
monomer 1 * 43000, SDS-PAGE Streptococcus pneumoniae
monomer 1 * 43000, SDS-PAGE Streptococcus pyogenes
monomer 1 * 46079, calculated from sequence Streptococcus pneumoniae
monomer 1 * 46100, calculated from sequence Streptococcus pyogenes
monomer 1 * 46587, calculated from sequence Streptococcus agalactiae

Synonyms

Synonyms Comment Organism
SagUGL
-
 Streptococcus agalactiae
SpnUGL
-
 Streptococcus pneumoniae
SpyUGL
-
 Streptococcus pyogenes

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Streptococcus agalactiae
30
-
 assay at Streptococcus pneumoniae
30
-
 assay at Streptococcus pyogenes
37
-
-
 Streptococcus agalactiae
37
-
-
 Streptococcus pneumoniae
37
-
-
 Streptococcus pyogenes

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
45
-
 20 mM Tris-HCl (pH 7.5), 5 min, 50% loss of activity Streptococcus agalactiae
45
-
 20 mM Tris-HCl (pH 7.5), 5 min, 50% loss of activity Streptococcus pneumoniae
45
-
 20 mM Tris-HCl (pH 7.5), 5 min, 50% loss of activity Streptococcus pyogenes

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.3
-
 beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S pH 7.5, 30°C, wild-type enzyme Streptococcus pneumoniae
4
-
 beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S pH 7.5, 30°C, wild-type enzyme Streptococcus pyogenes
24
-
 beta-D-4-deoxy-DELTA4,5-GlcAp-(1->3)-beta-D-GalNAc6S pH 7.5, 30°C, wild-type enzyme Streptococcus agalactiae

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.5
-
 in 20 mM potassium phosphate Streptococcus agalactiae
5.5
-
 in 20 mM potassium phosphate Streptococcus pneumoniae
5.5
-
 in 20 mM potassium phosphate Streptococcus pyogenes
7.5
-
 assay at Streptococcus agalactiae
7.5
-
 assay at Streptococcus pneumoniae
7.5
-
 assay at Streptococcus pyogenes

Expression

Organism Comment Expression
Streptococcus agalactiae constitutively expressed, expression level increases in presence of glycosaminoglycan up