Literature summary for 2.7.7.7 extracted from

Sherrer, S.M.; Maxwell, B.A.; Pack, L.R.; Fiala, K.A.; Fowler, J.D.; Zhang, J.; Suo, Z.
Identification of an unfolding intermediate for a DNA lesion bypass polymerase (2012), Chem. Res. Toxicol., 25, 1531-1540.

Search for more literature for 2.7.7.7

Cloned(Commentary)

Cloned (Comment)	Organism
wild-type Dpo4 is fused to a C-terminal His6-tag and overexpressed in Escherichia coli	Saccharolobus solfataricus

Organism

Organism	UniProt	Comment	Textmining
Saccharolobus solfataricus	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Saccharolobus solfataricus

Synonyms

Synonyms	Comment	Organism
DNA polymerase IV	-	Saccharolobus solfataricus
Dpo4	-	Saccharolobus solfataricus

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
additional information	-	wild-type enzyme exhibits a three-state cooperative unfolding profile, with an unfolding intermediate existing between the native and denatured states. The native state of wild-type Dpo4 persists from 26 to 87.5°C, and the unfolding intermediate is observed between 92 and 96.5 °C. Starting at 110 °C, the molar ellipticity approaches its highest value and does not change significantly in the temperature range of 110°C to 119°C. The Little Finger domain of the enzyme, which is only found in the Y-family DNA polymerases, is more thermostable than the polymerase core. The enzyme exhibits a three-state cooperative unfolding profile with an unfolding intermediate. The linker region between the Little Finger and Thumb domains of Dpo4 is a source of structural instability. Through site-directed mutagenesis, the interactions between the residues in the linker region and the Palm domain are identified to play a critical role in the formation of the unfolding intermediate. The secondary structure of Dpo4 is not altered when the temperature is increased from 26 to 87.5°C	Saccharolobus solfataricus
88	-	Tm-value of mutant enzyme E100A/E235A/R240A: 88.3°C	Saccharolobus solfataricus
89	-	Tm-value of mutant enzyme E100A: 88.9°C. Tm-value of mutant enzyme E100A/K148A: 89.4°C	Saccharolobus solfataricus
90	-	Tm-value of mutant enzyme E235A/R240A or mutant enzyme K148A/E235A/R240A: 90.3°C. Tm-value of mutant enzyme E100A/K148A/E235A/R240A: 90.0°C	Saccharolobus solfataricus
93	-	Tm-value of mutant enzyme K148A: 92.8°C	Saccharolobus solfataricus
97	-	Tm-value of mutant enzyme R240A: 96.5°C	Saccharolobus solfataricus
98	-	Tm-value of mutant enzyme P236A: 97.4°C	Saccharolobus solfataricus
100	-	Tm-value of wild-type enzyme: 99.6°C	Saccharolobus solfataricus

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Release 2023.1 (January 2023)