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Literature summary for 3.2.1.166 extracted from

  • Melo, C.M.; Tersariol, I.L.; Nader, H.B.; Pinhal, M.A.; Lima, M.A.
    Development of new methods for determining the heparanase enzymatic activity (2015), Carbohydr. Res., 412, 66-70 .
    View publication on PubMed
Search for more literature for 3.2.1.166

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of GST-tagged enzyme from vector pGEX-2TK-heparanase in Escherichia coli strain BL21 Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
heparan sulfate + H2O Homo sapiens
-
 ?
-
 ?

Organism

Organism UniProt Comment Textmining
Homo sapiens Q9Y251
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant GST-tagged enzyme from Escherichia coli strain BL21 by glutathione affinity chromatography Homo sapiens

Source Tissue

Source Tissue Comment Organism Textmining
MCF-7 cell
-
 Homo sapiens
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
fondaparinux + H2O
-
 Homo sapiens 2N-sulfo-6-O-sulfo-alpha-D-GlcN-(1->4)-beta-GlcA + a trisaccharide
-
 ?
heparan sulfate + H2O
-
 Homo sapiens ?
-
 ?
additional information development of two heparanase activity assays to cover a whole range of applications, overview. The first is a fast and easy method based on commercial homogenous substrate, fondaparinux and usage of a fluorescent redox marker, resazurin. Resazurin reduction reaction at pH 5.5 and 45°C. The second method is a quantitative assay based on biotinylated heparan sulfate that uses an easier technique to immobilize the substrate in a 96-well plate via protamine sulfate. Biotinylated heparan sulfate is immobilized. The enzymatic assay is performed using chimeric recombinant heparanase at different concentrations. In sequence, the immobilized biotinylated heparan sulfate that is not digested by recombinant heparanase is bound to streptavidin conjugated with europium. Fluorescence is measured using a time-resolved fluorometer. Both methods have high sensitivity and can be used to detect heparanase activity Homo sapiens ?
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 ?

Synonyms

Synonyms Comment Organism
endo-beta-glucuronidase
-
 Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
-
 assay at Homo sapiens

General Information

General Information Comment Organism
physiological function heparanase is a mammalian endo-beta-glucuronidase with importance in various pathological, e.g. carcinogenesis, and non-pathological events. Despite tumor development heparanase can also modulate inflammatory processes, scarring, tissue repair, and tissue regeneration Homo sapiens