Cloning, expression, and structure analysis of carbamate kinase-like carbamoyl phosphate synthetase from Pyrococcus abyssi
Purcarea, C.; Hervé, G.; Cunin, R.; Evans, D.R. (2001). Cloning, expression, and structure analysis of carbamate kinase-like carbamoyl phosphate synthetase from Pyrococcus abyssi. Extremophiles 5(4): 229-239. https://dx.doi.org/10.1007/s007920100201
In: Extremophiles. Springer: Tokyo. ISSN 1431-0651; e-ISSN 1433-4909, more
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| Keywords |
Pyrococcus abyssi Erauso, Reysenbach, Godfroy, Meunier, Crump, Partensky, Baross, Marteinsson, Barbier, Pace & Prieur, 1993 [WoRMS]
Marine/Coastal |
| Author keywords |
carbamate kinase; carbamoyl phosphate synthetase; Pyrococcus abyssi;evolution; archaea; hyperthermophily |
| Authors | | Top |
- Purcarea, C.
- Hervé, G.
- Cunin, R.
- Evans, D.R.
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| Abstract |
Pyrococcus abyssi, a hyperthermophilic archaeon found in the vicinity of deep-sea hydrothermal vents, grows optimally at temperatures around 100°C. Carbamoyl phosphate synthetase (CPSase) from this organism was cloned and sequenced. The active 34-kDa recombinant protein was overexpressed in Escherichia coli when the host cells were cotransformed with a plasmid encoding tRNA synthetases for low-frequency Escherichia coli codons. Sequence homology suggests that the tertiary structure of P. abyssi CPSase, resembling its counterpart in Pyrococcus furiosus, is closely related to the catabolic carbamate kinases and is very different from the larger mesophilic CPSases. P. furiosus CPSase and carbamate kinase form carbamoyl phosphate by phosphorylating carbamate produced spontaneously in solution from ammonia and bicarbonate. In contrast, P. abyssi CPSase has intrinsic bicarbonate-dependent ATPase activity, suggesting that the enzyme can catalyze the phosphorylation of the isosteric substrates carbamate and bicarbonate. |
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