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Literature summary extracted from

  • Alonso, A.; Terrados, G.; Picher, A.J.; Giraldo, R.; Blanco, L.; Larraga, V.
    An intrinsic 5'-deoxyribose-5-phosphate lyase activity in DNA polymerase beta from Leishmania infantum supports a role in DNA repair (2006), DNA repair, 5, 89-101.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.99.B1 expresses in Escherichia coli M15 (pREP4). The protein aggregates as inclusion bodies, solubilized in 5 M guanidinium hydrochloride and 20 mM inidazole-HCl, pH 8.0 and purified Leishmania infantum

Organism

EC Number Organism UniProt Comment Textmining
4.2.99.B1 Leishmania infantum Q9U6N3
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.99.B1 recombinant protein, expressed in Escherichia coli M15 (pREP4). The protein aggregates as inclusion bodies, solubilized in 5 M guanidinium hydrochloride and 20 mM imidazole-HCl, pH 8.0 and purified Leishmania infantum

Reaction

EC Number Reaction Comment Organism Reaction ID
4.2.99.B1 catalysis of the beta-elimination of the 5' deoxyribose-5-phosphate at an abasic site in DNA where a DNA-(apurinic or apyrimidinic site) lyase has already cleaved the C-O-P bond 3' to the apurinic or apyrimidinic site concerted reaction of dRP lyase and DNA polymerization of Li Polbeta on a uracil-containing DNA participating in single-nucleotide base excision repair (BER), beta-elimination reaction Leishmania infantum

Source Tissue

EC Number Source Tissue Comment Organism Textmining
4.2.99.B1 amastigote strain MCAN/ES/89/IPZ229/1/89, the enzyme activity varies with the life cycle of Leishmania infantum, being maximal in the intracellular amastigote phase Leishmania infantum
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4.2.99.B1 promastigote strain MHOM/FR/80/LEM75, the enzyme activity varies with the life cycle of Leishmania infantum, being maximal in the intracellular amastigote phase Leishmania infantum
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Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.2.99.B1 additional information
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the enzyme activity varies with the life cycle of Leishmania infantum, being maximal in the intracellular amastigote phase Leishmania infantum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.99.B1 34-bp-oligonucleotide containing uracil at position 16 labelled with 3'-end by terminal deoxynucleotidyltransferase using [alpha-32P]ddATP and annealed to its complementary oligonucleotide, and pretreated with human uracil-DNA glycosylase and AP endonuclease Leishmania infantum 19-bp-oligonucleotide + 2-deoxy-D-ribose 5-phosphate
-
?
4.2.99.B1 additional information the dRP removal proceeds through a beta-elimination, verified by addition of sodium borohydride forming a covalent protein-DNA complex by reduction of the Schiff base intermediate Leishmania infantum ?
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?

Subunits

EC Number Subunits Comment Organism
4.2.99.B1 More the 8-kDa domain of polymerase beta is responsible for single-stranded DNA binding, 5'-phosphate recognition, and dRP lyase activity, formed by the amino acid residues 1-91 of Li Polbeta. The residues Tyr39, Lys60, Lys68, Lys72, Glu75, and Glu84 play a role in the catalysis. Lys72, identified as the nucleophile in Polbeta is responsible for Schiff base forming during beta-elimination of the dRP moiety Leishmania infantum

Synonyms

EC Number Synonyms Comment Organism
4.2.99.B1 5'-deoxyribose-5-phosphate lyase
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Leishmania infantum
4.2.99.B1 dRP lyase
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Leishmania infantum
4.2.99.B1 Li polbeta
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Leishmania infantum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.2.99.B1 37
-
assay at Leishmania infantum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.2.99.B1 7.5
-
assay at Leishmania infantum