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Literature summary for 4.2.2.1 extracted from

  • Singh, S.K.; Malhotra, S.; Akhtar, M.S.
    Characterization of hyaluronic acid specific hyaluronate lyase (HylP) from Streptococcus pyogenes (2014), Biochimie, 102, 203-210.
    View publication on PubMed
Search for more literature for 4.2.2.1

Cloned(Commentary)

Cloned (Comment) Organism
DNA and amino acid sequence determination and analysis, recombinant expression of wild-type and truncation and point mutant enzymes in Escherichia coli strain BL21(DE3) using plasmid pSF49 Streptococcus pyogenes phage H4489A

Protein Variants

Protein Variants Comment Organism
D170T site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme Streptococcus pyogenes phage H4489A
additional information construction of truncated mutants hylp255e1113 and hylp390e1113, HylP enzyme without internal Gly-X-Y motif Streptococcus pyogenes phage H4489A
Q295E site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Streptococcus pyogenes phage H4489A
W19F site-directed mutagenesis, denaturation profile with guanidinium hydrochloride is similar to the wild-type enzyme Streptococcus pyogenes phage H4489A
Y182F site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme Streptococcus pyogenes phage H4489A
Y298F site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Streptococcus pyogenes phage H4489A

Inhibitors

Inhibitors Comment Organism Structure
Ca2+ interacts with the collagenous Gly-X-Y motif of the enzyme, activates up to 20 mM, but inhibits at higher concentrations, inactivation above 50 mM Streptococcus pyogenes phage H4489A
L-ascorbate noncompetitive inhibition, inhibition kinetics, overview. Residues involved in the binding of L-ascorbate are confined to HylP135-308 Streptococcus pyogenes phage H4489A

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information Michaelis-Menten kinetics Streptococcus pyogenes phage H4489A
0.285
-
 hyaluronic acid pH 6.0, 25°C, recombinant wild-type enzyme Streptococcus pyogenes phage H4489A
0.305
-
 hyaluronic acid pH 6.0, 25°C, recombinant mutant Y298F Streptococcus pyogenes phage H4489A
0.373
-
 hyaluronic acid pH 6.0, 25°C, recombinant mutant Q295E Streptococcus pyogenes phage H4489A

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ interacts with the collagenous Gly-X-Y motif of the enzyme, activates up to 20 mM, but inhibits at higher concentrations Streptococcus pyogenes phage H4489A

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
40000
-
 3 * 40000, SDS-PAGE Streptococcus pyogenes phage H4489A
120000
-
 recombinant enzyme, glutaraldehyde cross-linking Streptococcus pyogenes phage H4489A

Organism

Organism UniProt Comment Textmining
Streptococcus pyogenes phage H4489A P15316
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli Streptococcus pyogenes phage H4489A

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
hyaluronic acid the enzyme is a hyaluronic acid specific hyaluronate lyase showing ability to degrade hyaluronate to unsaturated disaccharide units Streptococcus pyogenes phage H4489A ?
-
 ?

Subunits

Subunits Comment Organism
More extended trimeric conformation, homology modeling using the crystal structure of HylP1 from Streptococcus pyogenes SF370 (PDB ID-2C3F) as a template, domain structure, overview Streptococcus pyogenes phage H4489A
trimer 3 * 40000, SDS-PAGE Streptococcus pyogenes phage H4489A

Synonyms

Synonyms Comment Organism
HylP
-
 Streptococcus pyogenes phage H4489A

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Streptococcus pyogenes phage H4489A

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.76
-
 hyaluronic acid pH 6.0, 25°C, recombinant mutant Q295E Streptococcus pyogenes phage H4489A
4.29
-
 hyaluronic acid pH 6.0, 25°C, recombinant mutant Y298F Streptococcus pyogenes phage H4489A
5.65
-
 hyaluronic acid pH 6.0, 25°C, recombinant wild-type enzyme Streptococcus pyogenes phage H4489A

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
 assay at Streptococcus pyogenes phage H4489A

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.7
-
 pH 6.0, 25°C, recombinant wild-type enzyme Streptococcus pyogenes phage H4489A L-ascorbate

General Information

General Information Comment Organism
malfunction the enzyme activity is considerably reduced with mutation in the second pocket consisting of Glu295 and Tyr298 Streptococcus pyogenes phage H4489A
additional information The primary catalytic residues of the enzyme seem to be in the first pocket consisting of Asp170 and Tyr182. Catalytic residues span between the regions containing 135-308 amino acids where both the catalytic pocket has a prominent positively charged residue, structure overview. The collagenous Gly-X-Y motif of the enzyme influences stability and interact with calcium ions suggesting its role in the enzyme regulation Streptococcus pyogenes phage H4489A
physiological function hyaluronate lyase is proposed to be one of the key bacteriophage-encoded virulence factors. The Gly-X-Y motif of HylP has a regulatory role for enzyme function Streptococcus pyogenes phage H4489A