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Globin-like proteins in Caenorhabditis elegans: in vivo localization, ligand binding and structural properties

BACKGROUND: The genome of the nematode Caenorhabditis elegans contains more than 30 putative globin genes that all are transcribed. Although their translated amino acid sequences fit the globin fold, a variety of amino-acid substitutions and extensions generate a wide structural diversity among the...

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Autores principales: Geuens, Eva, Hoogewijs, David, Nardini, Marco, Vinck, Evi, Pesce, Alessandra, Kiger, Laurent, Fago, Angela, Tilleman, Lesley, De Henau, Sasha, Marden, Michael C, Weber, Roy E, Van Doorslaer, Sabine, Vanfleteren, Jacques, Moens, Luc, Bolognesi, Martino, Dewilde, Sylvia
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2867796/
https://www.ncbi.nlm.nih.gov/pubmed/20361867
http://dx.doi.org/10.1186/1471-2091-11-17
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author Geuens, Eva
Hoogewijs, David
Nardini, Marco
Vinck, Evi
Pesce, Alessandra
Kiger, Laurent
Fago, Angela
Tilleman, Lesley
De Henau, Sasha
Marden, Michael C
Weber, Roy E
Van Doorslaer, Sabine
Vanfleteren, Jacques
Moens, Luc
Bolognesi, Martino
Dewilde, Sylvia
author_facet Geuens, Eva
Hoogewijs, David
Nardini, Marco
Vinck, Evi
Pesce, Alessandra
Kiger, Laurent
Fago, Angela
Tilleman, Lesley
De Henau, Sasha
Marden, Michael C
Weber, Roy E
Van Doorslaer, Sabine
Vanfleteren, Jacques
Moens, Luc
Bolognesi, Martino
Dewilde, Sylvia
author_sort Geuens, Eva
collection PubMed
description BACKGROUND: The genome of the nematode Caenorhabditis elegans contains more than 30 putative globin genes that all are transcribed. Although their translated amino acid sequences fit the globin fold, a variety of amino-acid substitutions and extensions generate a wide structural diversity among the putative globins. No information is available on the physicochemical properties and the in vivo expression. RESULTS: We expressed the globins in a bacterial system, characterized the purified proteins by optical and resonance Raman spectroscopy, measured the kinetics and equilibria of O(2 )binding and determined the crystal structure of GLB-1* (CysGH2 → Ser mutant). Furthermore, we studied the expression patterns of glb-1 (ZK637.13) and glb-26 (T22C1.2) in the worms using green fluorescent protein technology and measured alterations of their transcript abundances under hypoxic conditions.GLB-1* displays the classical three-over-three α-helical sandwich of vertebrate globins, assembled in a homodimer associated through facing E- and F-helices. Within the heme pocket the dioxygen molecule is stabilized by a hydrogen bonded network including TyrB10 and GlnE7.GLB-1 exhibits high ligand affinity, which is, however, lower than in other globins with the same distal TyrB10-GlnE7 amino-acid pair. In the absence of external ligands, the heme ferrous iron of GLB-26 is strongly hexacoordinated with HisE7, which could explain its extremely low affinity for CO. This globin oxidizes instantly to the ferric form in the presence of oxygen and is therefore incapable of reversible oxygen binding. CONCLUSION: The presented data indicate that GLB-1 and GLB-26 belong to two functionally-different globin classes.
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spelling pubmed-28677962010-05-12 Globin-like proteins in Caenorhabditis elegans: in vivo localization, ligand binding and structural properties Geuens, Eva Hoogewijs, David Nardini, Marco Vinck, Evi Pesce, Alessandra Kiger, Laurent Fago, Angela Tilleman, Lesley De Henau, Sasha Marden, Michael C Weber, Roy E Van Doorslaer, Sabine Vanfleteren, Jacques Moens, Luc Bolognesi, Martino Dewilde, Sylvia BMC Biochem Research article BACKGROUND: The genome of the nematode Caenorhabditis elegans contains more than 30 putative globin genes that all are transcribed. Although their translated amino acid sequences fit the globin fold, a variety of amino-acid substitutions and extensions generate a wide structural diversity among the putative globins. No information is available on the physicochemical properties and the in vivo expression. RESULTS: We expressed the globins in a bacterial system, characterized the purified proteins by optical and resonance Raman spectroscopy, measured the kinetics and equilibria of O(2 )binding and determined the crystal structure of GLB-1* (CysGH2 → Ser mutant). Furthermore, we studied the expression patterns of glb-1 (ZK637.13) and glb-26 (T22C1.2) in the worms using green fluorescent protein technology and measured alterations of their transcript abundances under hypoxic conditions.GLB-1* displays the classical three-over-three α-helical sandwich of vertebrate globins, assembled in a homodimer associated through facing E- and F-helices. Within the heme pocket the dioxygen molecule is stabilized by a hydrogen bonded network including TyrB10 and GlnE7.GLB-1 exhibits high ligand affinity, which is, however, lower than in other globins with the same distal TyrB10-GlnE7 amino-acid pair. In the absence of external ligands, the heme ferrous iron of GLB-26 is strongly hexacoordinated with HisE7, which could explain its extremely low affinity for CO. This globin oxidizes instantly to the ferric form in the presence of oxygen and is therefore incapable of reversible oxygen binding. CONCLUSION: The presented data indicate that GLB-1 and GLB-26 belong to two functionally-different globin classes. BioMed Central 2010-04-02 /pmc/articles/PMC2867796/ /pubmed/20361867 http://dx.doi.org/10.1186/1471-2091-11-17 Text en Copyright ©2010 Geuens et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research article
Geuens, Eva
Hoogewijs, David
Nardini, Marco
Vinck, Evi
Pesce, Alessandra
Kiger, Laurent
Fago, Angela
Tilleman, Lesley
De Henau, Sasha
Marden, Michael C
Weber, Roy E
Van Doorslaer, Sabine
Vanfleteren, Jacques
Moens, Luc
Bolognesi, Martino
Dewilde, Sylvia
Globin-like proteins in Caenorhabditis elegans: in vivo localization, ligand binding and structural properties
title Globin-like proteins in Caenorhabditis elegans: in vivo localization, ligand binding and structural properties
title_full Globin-like proteins in Caenorhabditis elegans: in vivo localization, ligand binding and structural properties
title_fullStr Globin-like proteins in Caenorhabditis elegans: in vivo localization, ligand binding and structural properties
title_full_unstemmed Globin-like proteins in Caenorhabditis elegans: in vivo localization, ligand binding and structural properties
title_short Globin-like proteins in Caenorhabditis elegans: in vivo localization, ligand binding and structural properties
title_sort globin-like proteins in caenorhabditis elegans: in vivo localization, ligand binding and structural properties
topic Research article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2867796/
https://www.ncbi.nlm.nih.gov/pubmed/20361867
http://dx.doi.org/10.1186/1471-2091-11-17
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