Engineered chimeras reveal the structural basis of hexacoordination in globins: A case study of neuroglobin and myoglobin
Background Myoglobin (Mb) and neuroglobin (Ngb) are representative members of pentacoordinated and bis-histidyl, hexacoordinated globins. In spite of their low sequence identity, they show surprisingly similar three-dimensional folds. The ability of Ngb to form a hexacoordinated bis-histidyl complex...
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03044165_v1850_n1_p169_Boron http://hdl.handle.net/20.500.12110/paper_03044165_v1850_n1_p169_Boron |
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paper:paper_03044165_v1850_n1_p169_Boron2023-06-08T15:29:53Z Engineered chimeras reveal the structural basis of hexacoordination in globins: A case study of neuroglobin and myoglobin Capece, Luciana Wetzler, Diana E. Martí, Marcelo Adrián Estrin, Dario Ariel Nadra, Alejandro Daniel Bis-histidyl coordination Flash photolysis Molecular dynamics Myoglobin Neuroglobin carbon monoxide chimeric protein myoglobin neuroglobin globin heme hybrid protein ligand myoglobin nerve protein neuroglobin protein binding Article binding kinetics CD region dissociation hexacoordination molecular dynamics photolysis protein engineering protein structure spectroscopy amino acid sequence animal chemistry genetics human metabolism molecular genetics protein tertiary structure regulatory sequence sequence homology spectrophotometry Amino Acid Sequence Animals Globins Heme Humans Ligands Molecular Dynamics Simulation Molecular Sequence Data Myoglobin Nerve Tissue Proteins Protein Binding Protein Engineering Protein Structure, Tertiary Recombinant Fusion Proteins Regulatory Sequences, Nucleic Acid Sequence Homology, Amino Acid Spectrophotometry Background Myoglobin (Mb) and neuroglobin (Ngb) are representative members of pentacoordinated and bis-histidyl, hexacoordinated globins. In spite of their low sequence identity, they show surprisingly similar three-dimensional folds. The ability of Ngb to form a hexacoordinated bis-histidyl complex with the distal HisE7 has a strong impact on ligand affinity. The factors governing such different behaviors have not been completely understood yet, even though they are extremely relevant to establish structure-function relationships within the globin superfamily. Methods In this work we generated chimeric proteins by swapping a previously identified regulatory segment - the CD region - and evaluated comparatively the structural and functional properties of the resulting proteins by molecular dynamics simulations, and spectroscopic and kinetic investigations. Results Our results show that chimeric proteins display heme coordination properties displaced towards those expected for the corresponding CD region. In particular, in the absence of exogenous ligands, chimeric Mb is found as a partially hexacoordinated bis-histidyl species, whereas chimeric Ngb shows a lower equilibrium constant for forming a hexacoordinated bis-histidyl species. Conclusions While these results confirm the regulatory role of the CD region for bis-histidyl hexacoordination, they also suggest that additional sources contribute to fine tune the equilibrium. General significance Globins constitute a ubiquitous group of heme proteins widely found in all kingdoms of life. These findings raise challenging questions regarding the structure-function relationships in these proteins, as bis-histidyl hexacoordination emerges as a novel regulatory mechanism of the physiological function of globins. ©2014 Elsevier B.V. All rights reserved. Fil:Capece, L. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Wetzler, D.E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Marti, M.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Estrin, D.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Nadra, A.D. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2015 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03044165_v1850_n1_p169_Boron http://hdl.handle.net/20.500.12110/paper_03044165_v1850_n1_p169_Boron |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Bis-histidyl coordination Flash photolysis Molecular dynamics Myoglobin Neuroglobin carbon monoxide chimeric protein myoglobin neuroglobin globin heme hybrid protein ligand myoglobin nerve protein neuroglobin protein binding Article binding kinetics CD region dissociation hexacoordination molecular dynamics photolysis protein engineering protein structure spectroscopy amino acid sequence animal chemistry genetics human metabolism molecular genetics protein tertiary structure regulatory sequence sequence homology spectrophotometry Amino Acid Sequence Animals Globins Heme Humans Ligands Molecular Dynamics Simulation Molecular Sequence Data Myoglobin Nerve Tissue Proteins Protein Binding Protein Engineering Protein Structure, Tertiary Recombinant Fusion Proteins Regulatory Sequences, Nucleic Acid Sequence Homology, Amino Acid Spectrophotometry |
| spellingShingle |
Bis-histidyl coordination Flash photolysis Molecular dynamics Myoglobin Neuroglobin carbon monoxide chimeric protein myoglobin neuroglobin globin heme hybrid protein ligand myoglobin nerve protein neuroglobin protein binding Article binding kinetics CD region dissociation hexacoordination molecular dynamics photolysis protein engineering protein structure spectroscopy amino acid sequence animal chemistry genetics human metabolism molecular genetics protein tertiary structure regulatory sequence sequence homology spectrophotometry Amino Acid Sequence Animals Globins Heme Humans Ligands Molecular Dynamics Simulation Molecular Sequence Data Myoglobin Nerve Tissue Proteins Protein Binding Protein Engineering Protein Structure, Tertiary Recombinant Fusion Proteins Regulatory Sequences, Nucleic Acid Sequence Homology, Amino Acid Spectrophotometry Capece, Luciana Wetzler, Diana E. Martí, Marcelo Adrián Estrin, Dario Ariel Nadra, Alejandro Daniel Engineered chimeras reveal the structural basis of hexacoordination in globins: A case study of neuroglobin and myoglobin |
| topic_facet |
Bis-histidyl coordination Flash photolysis Molecular dynamics Myoglobin Neuroglobin carbon monoxide chimeric protein myoglobin neuroglobin globin heme hybrid protein ligand myoglobin nerve protein neuroglobin protein binding Article binding kinetics CD region dissociation hexacoordination molecular dynamics photolysis protein engineering protein structure spectroscopy amino acid sequence animal chemistry genetics human metabolism molecular genetics protein tertiary structure regulatory sequence sequence homology spectrophotometry Amino Acid Sequence Animals Globins Heme Humans Ligands Molecular Dynamics Simulation Molecular Sequence Data Myoglobin Nerve Tissue Proteins Protein Binding Protein Engineering Protein Structure, Tertiary Recombinant Fusion Proteins Regulatory Sequences, Nucleic Acid Sequence Homology, Amino Acid Spectrophotometry |
| description |
Background Myoglobin (Mb) and neuroglobin (Ngb) are representative members of pentacoordinated and bis-histidyl, hexacoordinated globins. In spite of their low sequence identity, they show surprisingly similar three-dimensional folds. The ability of Ngb to form a hexacoordinated bis-histidyl complex with the distal HisE7 has a strong impact on ligand affinity. The factors governing such different behaviors have not been completely understood yet, even though they are extremely relevant to establish structure-function relationships within the globin superfamily. Methods In this work we generated chimeric proteins by swapping a previously identified regulatory segment - the CD region - and evaluated comparatively the structural and functional properties of the resulting proteins by molecular dynamics simulations, and spectroscopic and kinetic investigations. Results Our results show that chimeric proteins display heme coordination properties displaced towards those expected for the corresponding CD region. In particular, in the absence of exogenous ligands, chimeric Mb is found as a partially hexacoordinated bis-histidyl species, whereas chimeric Ngb shows a lower equilibrium constant for forming a hexacoordinated bis-histidyl species. Conclusions While these results confirm the regulatory role of the CD region for bis-histidyl hexacoordination, they also suggest that additional sources contribute to fine tune the equilibrium. General significance Globins constitute a ubiquitous group of heme proteins widely found in all kingdoms of life. These findings raise challenging questions regarding the structure-function relationships in these proteins, as bis-histidyl hexacoordination emerges as a novel regulatory mechanism of the physiological function of globins. ©2014 Elsevier B.V. All rights reserved. |
| author |
Capece, Luciana Wetzler, Diana E. Martí, Marcelo Adrián Estrin, Dario Ariel Nadra, Alejandro Daniel |
| author_facet |
Capece, Luciana Wetzler, Diana E. Martí, Marcelo Adrián Estrin, Dario Ariel Nadra, Alejandro Daniel |
| author_sort |
Capece, Luciana |
| title |
Engineered chimeras reveal the structural basis of hexacoordination in globins: A case study of neuroglobin and myoglobin |
| title_short |
Engineered chimeras reveal the structural basis of hexacoordination in globins: A case study of neuroglobin and myoglobin |
| title_full |
Engineered chimeras reveal the structural basis of hexacoordination in globins: A case study of neuroglobin and myoglobin |
| title_fullStr |
Engineered chimeras reveal the structural basis of hexacoordination in globins: A case study of neuroglobin and myoglobin |
| title_full_unstemmed |
Engineered chimeras reveal the structural basis of hexacoordination in globins: A case study of neuroglobin and myoglobin |
| title_sort |
engineered chimeras reveal the structural basis of hexacoordination in globins: a case study of neuroglobin and myoglobin |
| publishDate |
2015 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03044165_v1850_n1_p169_Boron http://hdl.handle.net/20.500.12110/paper_03044165_v1850_n1_p169_Boron |
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