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Literature summary for 4.2.99.B1 extracted from

  • Saxowsky, T.T.; Matsumoto, Y.; Englund, P.T.
    The mitochondrial DNA polymerase beta from Crithdia fasciculata has a 5'-deoxyribose phosphate (dRP) lyase activity but is deficient in the release of dRP (2002), J. Biol. Chem., 277, 37201-37206.
    View publication on PubMed
Search for more literature for 4.2.99.B1

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) via electroporation, lacking the 9-amino acid mitochondrial targeting signal sequence Crithidia fasciculata

Protein Variants

Protein Variants Comment Organism
K72A PCR-based site-directed mutagenesis, deficient in forming a Schiff base intermediate and accumulating an enzyme dRP intermediate without releasing free dRP Crithidia fasciculata

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00045
-
 28-mer deoxyoligonucleotide with a 5' uracil residue pretreated with Eschericia coli uracil N-glycosylase, with concentrations 0.1-2.0 microMol, kcat/Km: 0.24 microMol/min Crithidia fasciculata

Localization

Localization Comment Organism GeneOntology No. Textmining
kinetoplast containing unusual DNA, i.e. kinetoplast DNA or kDNA Crithidia fasciculata 20023
-
mitochondrion
-
 Crithidia fasciculata 5739
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
43000
-
 about, wild-type mitochondrial pol beta and K72A mutant Crithidia fasciculata
45000
-
 1 * 45000, SDS-PAGE, wild-type mitochondrial pol beta and K72A mutant Crithidia fasciculata

Organism

Organism UniProt Comment Textmining
Crithidia fasciculata
-
-
-

Purification (Commentary)

Purification (Comment) Organism
native and recombinant protein Crithidia fasciculata

Reaction

Reaction Comment Organism Reaction ID
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 the mitochondrial DNA polymerase in the parasite Crithidia fasciculata exhibits dRP lyase but shows deficiency in the release of dRP from the enzyme following its cleavage from the DNA Crithidia fasciculata
a

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
28-mer deoxyoligonucleotide with a 5' uracil residue pretreated with Eschericia coli uracil N-glycosylase Crithidia fasciculata ?
-
 ?

Subunits

Subunits Comment Organism
monomer 1 * 45000, SDS-PAGE, wild-type mitochondrial pol beta and K72A mutant Crithidia fasciculata
More alignment with the rat nuclear enzyme shows that the mitochondrial pol beta contains the essential conserved amino acids Lys35, Lys60, Lys68, and Lys72 Crithidia fasciculata

Synonyms

Synonyms Comment Organism
5'-deoxyribose phosphate lyase
-
 Crithidia fasciculata
dRP lyase
-
 Crithidia fasciculata

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Crithidia fasciculata

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0018
-
 28-mer deoxyoligonucleotide with a 5' uracil residue pretreated with Eschericia coli uracil N-glycosylase, with concentrations 0.1-2.0 microMol, kcat/Km: 0.24 microMol/min Crithidia fasciculata

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.6
-
 assay at Crithidia fasciculata