2011
DOI: 10.1002/cbic.201100145
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Role of Disulfide Bridges in Archaeal Family‐B DNA Polymerases

Abstract: The family-B DNA polymerases obtained from the order Thermococcales, for example, Pyrococcus furiosus (Pfu-Pol) are commonly used in the polymerase chain reaction (PCR) because of their high thermostability and low error rates. Most of these polymerases contain four cysteines, arranged as two disulfide bridges. With Pfu-Pol C429-C443 forms one of the disulfides (DB1) and C507-C510 (DB2) the other. Although the disulfides are well conserved in the enzymes from the hyperthermophilic Thermococcales, they are less… Show more

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Cited by 4 publications
(4 citation statements)
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References 47 publications
(15 reference statements)
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“…As postulated earlier it is probable that a second melting event for Pfu-Pol takes place above 100°C; therefore, only the first observed transitions have been used for comparing unfolding. The T m of 93.2°C is slightly higher than that of 89.7°C observed earlier for Pfu-Pol in 2 M GuHCl (Killelea and Connolly, 2011). Such variation may be accounted for by the different NaCl concentrations used, 400 mM here, 200 mM previously.…”
Section: Resultscontrasting
confidence: 60%
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“…As postulated earlier it is probable that a second melting event for Pfu-Pol takes place above 100°C; therefore, only the first observed transitions have been used for comparing unfolding. The T m of 93.2°C is slightly higher than that of 89.7°C observed earlier for Pfu-Pol in 2 M GuHCl (Killelea and Connolly, 2011). Such variation may be accounted for by the different NaCl concentrations used, 400 mM here, 200 mM previously.…”
Section: Resultscontrasting
confidence: 60%
“…Pyrococcus furiosus grows optimally at 100°C and can survive at 110°C, whereas Thermococcus kodakarensis grows best at 85°C and can tolerate 94°C (Fiala and Stetter, 1986; Borges et al, 2010). Our group earlier measured the thermostability of Pfu-Pol using differential scanning fluorimetry (DSF), where the protein is subject to a steady increase in temperature in the presence of the dye SYPRO orange (Killelea and Connolly, 2011). As thermally-induced unfolding takes place the dye binds to exposed hydrophobic regions, resulting in an increase in fluorescence that can be measured using a real time PCR apparatus.…”
Section: Resultsmentioning
confidence: 99%
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“…Noronha et all [6] point out that, as a way to achieve certain goals, decentralization was the only organizational guideline of the SUS that did not go against the so-called 'neo-liberal ideas' of strengthening the right to health and opposing the expansion of size of the state and…”
Section: Introductionmentioning
confidence: 99%