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Characterization of a globin-coupled oxygen sensor with a gene-regulating function

(2007) JOURNAL OF BIOLOGICAL CHEMISTRY. 282(52). p.37325-37340
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Abstract
Globin-coupled sensors (GCSs) are multiple-domain transducers, consisting of a regulatory globin-like heme-binding domain and a linked transducer domain(s). GCSs have been described in both Archaea and bacteria. They are generally assumed to bind O-2 (and perhaps other gaseous ligands) and to transmit a conformational change signal through the transducer domain in response to fluctuating O2 levels. In this study, the heme-binding domain, AvGReg178, and the full protein, AvGReg of the Azotobacter vinelandii GCS, were cloned, expressed, and purified. After purification, the heme iron of AvGReg178 was found to bind O2. This form was stable over many hours. In contrast, the predominant presence of a bis-histidinecoordinate heme in ferric AvGReg was revealed. Differences in the hemepocket structure were also observed for the deoxygenated ferrous state of these proteins. The spectra showed that the deoxygenated ferrous derivatives of AvGReg178 and AvGReg are characterized by a penta-coordinate and hexa-coordinate heme iron, respectively. O2 binding isotherms indicate that AvGReg178 and AvGReg show a high affinity for O-2 with P-50 values at 20 degrees C of 0.04 and 0.15 torr, respectively. Kinetics of CO binding indicate that AvGReg178 carbonylation conforms to a monophasic process, comparable with that of myoglobin, whereas AvGReg carbonylation conforms to a three-phasic reaction, as observed for several proteins with bis-histidine heme iron coordination. Besides sensing ligands, in vitro data suggest that AvGReg(178) may have a role in O-2-mediated NO-detoxification, yielding metAvGReg(178) and nitrate.
Keywords
HEME-BASED SENSORS, ELECTRON-PARAMAGNETIC-RES, TRUNCATED HEMOGLOBIN-O, HYDROGEN-BOND NETWORK, CYCLIC DI-GMP, NITRIC-OXIDE, LIGAND-BINDING, BACILLUS-SUBTILIS, RESONANCE RAMAN, MYCOBACTERIUM-LEPRAE

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Citation

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MLA
Thijs, Liesbet, Evi Vinck, Alessandro Bolli, et al. “Characterization of a Globin-coupled Oxygen Sensor with a Gene-regulating Function.” JOURNAL OF BIOLOGICAL CHEMISTRY 282.52 (2007): 37325–37340. Print.
APA
Thijs, L., Vinck, E., Bolli, A., Trandafir, F., Wan, X., Hoogewijs, D., Coletta, M., et al. (2007). Characterization of a globin-coupled oxygen sensor with a gene-regulating function. JOURNAL OF BIOLOGICAL CHEMISTRY, 282(52), 37325–37340.
Chicago author-date
Thijs, Liesbet, Evi Vinck, Alessandro Bolli, Florin Trandafir, Xuehua Wan, David Hoogewijs, Massimiliano Coletta, et al. 2007. “Characterization of a Globin-coupled Oxygen Sensor with a Gene-regulating Function.” Journal of Biological Chemistry 282 (52): 37325–37340.
Chicago author-date (all authors)
Thijs, Liesbet, Evi Vinck, Alessandro Bolli, Florin Trandafir, Xuehua Wan, David Hoogewijs, Massimiliano Coletta, Angela Fago, Roy E Weber, Sabine Van Doorslaer, Paolo Ascenzi, Maqsudul Alam, Luc Moens, and Sylvia Dewilde. 2007. “Characterization of a Globin-coupled Oxygen Sensor with a Gene-regulating Function.” Journal of Biological Chemistry 282 (52): 37325–37340.
Vancouver
1.
Thijs L, Vinck E, Bolli A, Trandafir F, Wan X, Hoogewijs D, et al. Characterization of a globin-coupled oxygen sensor with a gene-regulating function. JOURNAL OF BIOLOGICAL CHEMISTRY. 2007;282(52):37325–40.
IEEE
[1]
L. Thijs et al., “Characterization of a globin-coupled oxygen sensor with a gene-regulating function,” JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 282, no. 52, pp. 37325–37340, 2007.
@article{419659,
  abstract     = {Globin-coupled sensors (GCSs) are multiple-domain transducers, consisting of a regulatory globin-like heme-binding domain and a linked transducer domain(s). GCSs have been described in both Archaea and bacteria. They are generally assumed to bind O-2 (and perhaps other gaseous ligands) and to transmit a conformational change signal through the transducer domain in response to fluctuating O2 levels. In this study, the heme-binding domain, AvGReg178, and the full protein, AvGReg of the Azotobacter vinelandii GCS, were cloned, expressed, and purified. After purification, the heme iron of AvGReg178 was found to bind O2. This form was stable over many hours. In contrast, the predominant presence of a bis-histidinecoordinate heme in ferric AvGReg was revealed. Differences in the hemepocket structure were also observed for the deoxygenated ferrous state of these proteins. The spectra showed that the deoxygenated ferrous derivatives of AvGReg178 and AvGReg are characterized by a penta-coordinate and hexa-coordinate heme iron, respectively. O2 binding isotherms indicate that AvGReg178 and AvGReg show a high affinity for O-2 with P-50 values at 20 degrees C of 0.04 and 0.15 torr, respectively. Kinetics of CO binding indicate that AvGReg178 carbonylation conforms to a monophasic process, comparable with that of myoglobin, whereas AvGReg carbonylation conforms to a three-phasic reaction, as observed for several proteins with bis-histidine heme iron coordination. Besides sensing ligands, in vitro data suggest that AvGReg(178) may have a role in O-2-mediated NO-detoxification, yielding metAvGReg(178) and nitrate.},
  author       = {Thijs, Liesbet and Vinck, Evi and Bolli, Alessandro and Trandafir, Florin and Wan, Xuehua and Hoogewijs, David and Coletta, Massimiliano and Fago, Angela and Weber, Roy E and Van Doorslaer, Sabine and Ascenzi, Paolo and Alam, Maqsudul and Moens, Luc and Dewilde, Sylvia},
  issn         = {0021-9258},
  journal      = {JOURNAL OF BIOLOGICAL CHEMISTRY},
  keywords     = {HEME-BASED SENSORS,ELECTRON-PARAMAGNETIC-RES,TRUNCATED HEMOGLOBIN-O,HYDROGEN-BOND NETWORK,CYCLIC DI-GMP,NITRIC-OXIDE,LIGAND-BINDING,BACILLUS-SUBTILIS,RESONANCE RAMAN,MYCOBACTERIUM-LEPRAE},
  language     = {eng},
  number       = {52},
  pages        = {37325--37340},
  title        = {Characterization of a globin-coupled oxygen sensor with a gene-regulating function},
  url          = {http://dx.doi.org/10.1074/jbc.M705541200},
  volume       = {282},
  year         = {2007},
}

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