Roles of glutamates and metal ions in a rationally designed nitric oxide reductase based on myoglobin
A structural and functional model of bacterial nitric oxide reductase (NOR) has been designed by introducing two glutamates (Glu) and three histidines (His) in sperm whale myoglobin. X-ray structural data indicate that the three His and one Glu (V68E) residues bind iron, mimicking the putative FeB s...
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ftpubmed:oai:pubmedcentral.nih.gov:2889330 2023-05-15T18:26:49+02:00 Roles of glutamates and metal ions in a rationally designed nitric oxide reductase based on myoglobin Lin, Ying-Wu Yeung, Natasha Gao, Yi-Gui Miner, Kyle D. Tian, Shiliang Robinson, Howard Lu, Yi 2010-05-11 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2889330 http://www.ncbi.nlm.nih.gov/pubmed/20421510 https://doi.org/10.1073/pnas.1000526107 en eng National Academy of Sciences http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2889330 http://www.ncbi.nlm.nih.gov/pubmed/20421510 http://dx.doi.org/10.1073/pnas.1000526107 Biological Sciences Text 2010 ftpubmed https://doi.org/10.1073/pnas.1000526107 2013-09-03T01:40:16Z A structural and functional model of bacterial nitric oxide reductase (NOR) has been designed by introducing two glutamates (Glu) and three histidines (His) in sperm whale myoglobin. X-ray structural data indicate that the three His and one Glu (V68E) residues bind iron, mimicking the putative FeB site in NOR, while the second Glu (I107E) interacts with a water molecule and forms a hydrogen bonding network in the designed protein. Unlike the first Glu (V68E), which lowered the heme reduction potential by ∼110 mV, the second Glu has little effect on the heme potential, suggesting that the negatively charged Glu has a different role in redox tuning. More importantly, introducing the second Glu resulted in a ∼100% increase in NOR activity, suggesting the importance of a hydrogen bonding network in facilitating proton delivery during NOR reactivity. In addition, EPR and X-ray structural studies indicate that the designed protein binds iron, copper, or zinc in the FeB site, each with different effects on the structures and NOR activities, suggesting that both redox activity and an intermediate five-coordinate heme-NO species are important for high NOR activity. The designed protein offers an excellent model for NOR and demonstrates the power of using designed proteins as a simpler and more well-defined system to address important chemical and biological issues. Text Sperm whale PubMed Central (PMC) Proceedings of the National Academy of Sciences 107 19 8581 8586 |
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Biological Sciences Lin, Ying-Wu Yeung, Natasha Gao, Yi-Gui Miner, Kyle D. Tian, Shiliang Robinson, Howard Lu, Yi Roles of glutamates and metal ions in a rationally designed nitric oxide reductase based on myoglobin |
topic_facet |
Biological Sciences |
description |
A structural and functional model of bacterial nitric oxide reductase (NOR) has been designed by introducing two glutamates (Glu) and three histidines (His) in sperm whale myoglobin. X-ray structural data indicate that the three His and one Glu (V68E) residues bind iron, mimicking the putative FeB site in NOR, while the second Glu (I107E) interacts with a water molecule and forms a hydrogen bonding network in the designed protein. Unlike the first Glu (V68E), which lowered the heme reduction potential by ∼110 mV, the second Glu has little effect on the heme potential, suggesting that the negatively charged Glu has a different role in redox tuning. More importantly, introducing the second Glu resulted in a ∼100% increase in NOR activity, suggesting the importance of a hydrogen bonding network in facilitating proton delivery during NOR reactivity. In addition, EPR and X-ray structural studies indicate that the designed protein binds iron, copper, or zinc in the FeB site, each with different effects on the structures and NOR activities, suggesting that both redox activity and an intermediate five-coordinate heme-NO species are important for high NOR activity. The designed protein offers an excellent model for NOR and demonstrates the power of using designed proteins as a simpler and more well-defined system to address important chemical and biological issues. |
format |
Text |
author |
Lin, Ying-Wu Yeung, Natasha Gao, Yi-Gui Miner, Kyle D. Tian, Shiliang Robinson, Howard Lu, Yi |
author_facet |
Lin, Ying-Wu Yeung, Natasha Gao, Yi-Gui Miner, Kyle D. Tian, Shiliang Robinson, Howard Lu, Yi |
author_sort |
Lin, Ying-Wu |
title |
Roles of glutamates and metal ions in a rationally designed nitric oxide reductase based on myoglobin |
title_short |
Roles of glutamates and metal ions in a rationally designed nitric oxide reductase based on myoglobin |
title_full |
Roles of glutamates and metal ions in a rationally designed nitric oxide reductase based on myoglobin |
title_fullStr |
Roles of glutamates and metal ions in a rationally designed nitric oxide reductase based on myoglobin |
title_full_unstemmed |
Roles of glutamates and metal ions in a rationally designed nitric oxide reductase based on myoglobin |
title_sort |
roles of glutamates and metal ions in a rationally designed nitric oxide reductase based on myoglobin |
publisher |
National Academy of Sciences |
publishDate |
2010 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2889330 http://www.ncbi.nlm.nih.gov/pubmed/20421510 https://doi.org/10.1073/pnas.1000526107 |
genre |
Sperm whale |
genre_facet |
Sperm whale |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2889330 http://www.ncbi.nlm.nih.gov/pubmed/20421510 http://dx.doi.org/10.1073/pnas.1000526107 |
op_doi |
https://doi.org/10.1073/pnas.1000526107 |
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Proceedings of the National Academy of Sciences |
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107 |
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19 |
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8581 |
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8586 |
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1766208782647427072 |