Flavohemoglobin Hmp, but not its individual domains, confers protection from respiratory inhibition by nitric oxide in Escherichia coli
Escherichia coli possesses a two-domain flavohemoglobin, Hmp, implicated in nitric oxide (NO) detoxification. To determine the contribution of each domain of Hmp toward NO detoxification, we genetically engineered the Hmp protein and separately expressed the heme (HD) and the flavin (FD) domains in...
| Main Authors: | , , , , , , , |
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| Format: | Journal Article |
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The American Society for Biochemistry and Molecular Biology Inc
2003
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| Online Access: | http://hdl.handle.net/20.500.11937/24960 |
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| author | Hernndez-Urza, E. Mills, C. White, Greg Contreras-Zentella, M. Escamilla, E. Vasudevan, S. Membrillo-Hernndez, J. Poole, R. |
| author_facet | Hernndez-Urza, E. Mills, C. White, Greg Contreras-Zentella, M. Escamilla, E. Vasudevan, S. Membrillo-Hernndez, J. Poole, R. |
| author_sort | Hernndez-Urza, E. |
| building | Curtin Institutional Repository |
| collection | Online Access |
| description | Escherichia coli possesses a two-domain flavohemoglobin, Hmp, implicated in nitric oxide (NO) detoxification. To determine the contribution of each domain of Hmp toward NO detoxification, we genetically engineered the Hmp protein and separately expressed the heme (HD) and the flavin (FD) domains in a defined hmp mutant. Expression of each domain was confirmed by Western blot analysis. CO-difference spectra showed that the HD of Hmp can bind CO, but the CO adduct showed a slightly blue-shifted peak. Overexpression of the HD resulted in an improvement of growth to a similar extent to that observed with the Vitreoscilla hemeonly globin Vgb, whereas the FD alone did not improve growth. Viability of the hmp mutant in the presence of lethal concentrations of sodium nitroprusside was increased(to 30% survival after 2 h in 5 mM sodium nitroprusside) by overexpressing Vgb or the HD. However, maximal protection was provided only by holo-Hmp (75% survival under the same conditions). Cellular respiration of the hmp mutant was instantaneously inhibited in the presence of 13.5 M NO but remained insensitive to NO inhibition when these cells overexpressed Hmp. When HD or FD was expressed separately, no significant protection was observed. By contrast, overexpression of Vgb provided partial protection from NO respiratory inhibition. Our results suggest that, despite the homology between the HD from Hmp and Vgb (45% identity), their roles seem to be quite distinct. |
| first_indexed | 2025-11-14T07:54:53Z |
| format | Journal Article |
| id | curtin-20.500.11937-24960 |
| institution | Curtin University Malaysia |
| institution_category | Local University |
| last_indexed | 2025-11-14T07:54:53Z |
| publishDate | 2003 |
| publisher | The American Society for Biochemistry and Molecular Biology Inc |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | curtin-20.500.11937-249602017-09-13T15:52:52Z Flavohemoglobin Hmp, but not its individual domains, confers protection from respiratory inhibition by nitric oxide in Escherichia coli Hernndez-Urza, E. Mills, C. White, Greg Contreras-Zentella, M. Escamilla, E. Vasudevan, S. Membrillo-Hernndez, J. Poole, R. dihydropteridine reductase E. coli flavohemoglobins NO Escherichia coli possesses a two-domain flavohemoglobin, Hmp, implicated in nitric oxide (NO) detoxification. To determine the contribution of each domain of Hmp toward NO detoxification, we genetically engineered the Hmp protein and separately expressed the heme (HD) and the flavin (FD) domains in a defined hmp mutant. Expression of each domain was confirmed by Western blot analysis. CO-difference spectra showed that the HD of Hmp can bind CO, but the CO adduct showed a slightly blue-shifted peak. Overexpression of the HD resulted in an improvement of growth to a similar extent to that observed with the Vitreoscilla hemeonly globin Vgb, whereas the FD alone did not improve growth. Viability of the hmp mutant in the presence of lethal concentrations of sodium nitroprusside was increased(to 30% survival after 2 h in 5 mM sodium nitroprusside) by overexpressing Vgb or the HD. However, maximal protection was provided only by holo-Hmp (75% survival under the same conditions). Cellular respiration of the hmp mutant was instantaneously inhibited in the presence of 13.5 M NO but remained insensitive to NO inhibition when these cells overexpressed Hmp. When HD or FD was expressed separately, no significant protection was observed. By contrast, overexpression of Vgb provided partial protection from NO respiratory inhibition. Our results suggest that, despite the homology between the HD from Hmp and Vgb (45% identity), their roles seem to be quite distinct. 2003 Journal Article http://hdl.handle.net/20.500.11937/24960 10.1074/jbc.M303629200 The American Society for Biochemistry and Molecular Biology Inc unknown |
| spellingShingle | dihydropteridine reductase E. coli flavohemoglobins NO Hernndez-Urza, E. Mills, C. White, Greg Contreras-Zentella, M. Escamilla, E. Vasudevan, S. Membrillo-Hernndez, J. Poole, R. Flavohemoglobin Hmp, but not its individual domains, confers protection from respiratory inhibition by nitric oxide in Escherichia coli |
| title | Flavohemoglobin Hmp, but not its individual domains, confers protection from respiratory inhibition by nitric oxide in Escherichia coli |
| title_full | Flavohemoglobin Hmp, but not its individual domains, confers protection from respiratory inhibition by nitric oxide in Escherichia coli |
| title_fullStr | Flavohemoglobin Hmp, but not its individual domains, confers protection from respiratory inhibition by nitric oxide in Escherichia coli |
| title_full_unstemmed | Flavohemoglobin Hmp, but not its individual domains, confers protection from respiratory inhibition by nitric oxide in Escherichia coli |
| title_short | Flavohemoglobin Hmp, but not its individual domains, confers protection from respiratory inhibition by nitric oxide in Escherichia coli |
| title_sort | flavohemoglobin hmp, but not its individual domains, confers protection from respiratory inhibition by nitric oxide in escherichia coli |
| topic | dihydropteridine reductase E. coli flavohemoglobins NO |
| url | http://hdl.handle.net/20.500.11937/24960 |