Literature summary for 4.2.99.B1 extracted from

Prasad, R.; Longley, M.J.; Sharief, F.S.; Hou, E.W.; Copeland, W.C.; Wilson, S.H.
Human DNA polymerase theta possesses 5'-dRP lyase activity and functions in single-nucleotide base excision repair in vitro (2009), Nucleic Acids Res., 37, 1868-1877.

Search for more literature for 4.2.99.B1

Cloned(Commentary)

Cloned (Comment)	Organism
the 98-kDa-fragment and the mutant D829N/E830Q enzyme are overexpressed in Escherichia coli BL21-CodonPlus-RP-cells	Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
D829N/E830Q	the polymerase inactive mutant D829N/E830Q retains full 5'-dRP lyase activity. Domain mapping of the 98-kDa enzyme reveals that the 5'-dRP lyase active site resides in a 24-kDa-domain	Homo sapiens

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.009	-	35-bp-deoxyoligonucleotide duplex containing a uracil residue at position 15	full-length 290-kDa Pol theta, pH 7.5, 37°C, kcat/Km: 0.047 microM/min	Homo sapiens
0.0097	-	35-bp-deoxyoligonucleotide duplex containing a uracil residue at position 15	98-kDa fragment of Pol theta, pH 7.5, 37°C, kcat/Km: 0.072 microM/min	Homo sapiens

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
additional information	-	polymerase theta is a ca. 300 kDa polypetide. The 98-kDa C-terminal region possesses both the DNA polymerase and the dRP lyase activity. The 5'-dRP lyase activity is independent of the polymerase activity. The polymerase inactive mutant D829N/E830Q retains full 5'-dRP lyase activity. Domain mapping of the 98-kDa enzyme reveals that the 5'-dRP lyase active site resides in a 24-kDa-domain	Homo sapiens

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant C-terminal 98-kDA-fragment and the mutant enzyme D829N/E830Q are purified to near homogeneity	Homo sapiens

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	enzyme is involved in single-nucleotide base excision DNA repair, SN-BER	Homo sapiens	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
34-bp-deoxyoligonucleotide duplex containing a uracil residue at position 16	preincised [32P] apurinic/apyrimidinic site containing DNA, pretreated with uracil DNA-glycosylase and AP endonuclease	Homo sapiens	? + 2-deoxy-D-ribose phosphate	the reaction products are separarted by electrophoresis	?
35-bp-deoxyoligonucleotide duplex containing a uracil residue at position 15	in vitro single-nucleotide base excision DNA repair assay, containing amongst others uracil DNA-glycosylase, AP endonuclease and ligase I	Homo sapiens	? + 2-deoxy-D-ribose phosphate	-	?

Subunits

Subunits	Comment	Organism
More	polymerase theta is a ca. 300 kDa polypetide. The 98-kDa C-terminal region possesses both the DNA polymerase and the dRP lyase activity. The 5'-dRP lyase activity is independent of the polymerase activity. The polymerase inactive mutant D829N/E830Q retains full 5'-dRP lyase activity. Domain mapping of the 98-kDa enzyme reveals that the 5'-dRP lyase active site resides in a 24-kDa-domain	Homo sapiens

Synonyms

Synonyms	Comment	Organism
5'-deoxyribose phosphate lyase	-	Homo sapiens
5'-dRP lyase	-	Homo sapiens
DNA polymerase theta	exhibits 5'-deoxyribose phosphate lyase activity	Homo sapiens
Pol theta	exhibits 5'-deoxyribose phosphate lyase activity	Homo sapiens

Temperature Optimum [°C]

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

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.007	-	35-bp-deoxyoligonucleotide duplex containing a uracil residue at position 15	full-length 290-kDa Pol theta, pH 7.5, 37°C, kcat/Km: 0.047 microM/min	Homo sapiens
0.01167	-	35-bp-deoxyoligonucleotide duplex containing a uracil residue at position 15	98-kDa fragment of Pol theta, pH 7.5, 37°C, kcat/Km: 0.072 microM/min	Homo sapiens

pH Optimum

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

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Release 2023.1 (January 2023)