Structural insights into the allosteric site of Arabidopsis NADP-malic enzyme 2: role of the second sphere residues in the regulatory signal transmission

Structure–function studies contribute to deciphering how small modifcations in the primary structure could introduce desirable characteristics into enzymes without afecting its overall functioning. Malic enzymes (ME) are ubiquitous and responsible for a wide variety of functions. The availability of...

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Autores principales: Gerrard Wheeler, Mariel Claudia, Arias, Cintia Lucía, Da Fonseca Rezende e Mello, Juliana, Cirauqui Diaz, Nuria, Rodrigues, Carlos Rangel, Drincovich, María Fabiana, Mendonça Teles de Souza, Alessandra, Alvarez, Clarisa Ester
Formato: article artículo publishedVersion
Lenguaje:Inglés
Publicado: Springer 2022
Materias:
Malic enzyme
Fumarate regulation
Structure–function
Acceso en línea:http://hdl.handle.net/2133/23339
http://hdl.handle.net/2133/23339
Aporte de:
Repositorio Hipermedial de la Universidad Nacional de Rosario (UNR) de Universidad Nacional de Rosario
id I15-R121-2133-23339
record_format dspace
institution Universidad Nacional de Rosario
institution_str I-15
repository_str R-121
collection Repositorio Hipermedial de la Universidad Nacional de Rosario (UNR)
language Inglés
orig_language_str_mv eng
topic Malic enzyme
Fumarate regulation
Structure–function
spellingShingle Malic enzyme
Fumarate regulation
Structure–function
Gerrard Wheeler, Mariel Claudia
Arias, Cintia Lucía
Da Fonseca Rezende e Mello, Juliana
Cirauqui Diaz, Nuria
Rodrigues, Carlos Rangel
Drincovich, María Fabiana
Mendonça Teles de Souza, Alessandra
Alvarez, Clarisa Ester
Structural insights into the allosteric site of Arabidopsis NADP-malic enzyme 2: role of the second sphere residues in the regulatory signal transmission
topic_facet Malic enzyme
Fumarate regulation
Structure–function
description Structure–function studies contribute to deciphering how small modifcations in the primary structure could introduce desirable characteristics into enzymes without afecting its overall functioning. Malic enzymes (ME) are ubiquitous and responsible for a wide variety of functions. The availability of a high number of ME crystal structures from diferent species facilitates comparisons between sequence and structure. Specifcally, the structural determinants necessary for fumarate allosteric regulation of ME has been of particular interest. NADP-ME2 from Arabidopsis thaliana exhibits a distinctive and complex regulation by fumarate, acting as an activator or an inhibitor according to substrate and efector concentrations. However, the 3D structure for this enzyme is not yet reported. In this work, we characterized the NADP-ME2 allosteric site by structural modeling, molecular docking, normal mode analysis and mutagenesis. The regulatory site model and its docking analysis suggested that other C4 acids including malate, NADP-ME2 substrate, could also ft into fumarate’s pocket. Besides, a non-conserved cluster of hydrophobic residues in the second sphere of the allosteric site was identifed. The substitution of one of those residues, L62, by a less fexible residue as tryptophan, resulted in a complete loss of fumarate activation and a reduction of substrate afnities for the active site. In addition, normal mode analysis indicated that conformational changes leading to the activation could originate in the region surrounding L62, extending through the allosteric site till the active site. Finally, the results in this work contribute to the understanding of structural determinants necessary for allosteric regulation providing new insights for enzyme optimization.
format article
artículo
publishedVersion
author Gerrard Wheeler, Mariel Claudia
Arias, Cintia Lucía
Da Fonseca Rezende e Mello, Juliana
Cirauqui Diaz, Nuria
Rodrigues, Carlos Rangel
Drincovich, María Fabiana
Mendonça Teles de Souza, Alessandra
Alvarez, Clarisa Ester
author_facet Gerrard Wheeler, Mariel Claudia
Arias, Cintia Lucía
Da Fonseca Rezende e Mello, Juliana
Cirauqui Diaz, Nuria
Rodrigues, Carlos Rangel
Drincovich, María Fabiana
Mendonça Teles de Souza, Alessandra
Alvarez, Clarisa Ester
author_sort Gerrard Wheeler, Mariel Claudia
title Structural insights into the allosteric site of Arabidopsis NADP-malic enzyme 2: role of the second sphere residues in the regulatory signal transmission
title_short Structural insights into the allosteric site of Arabidopsis NADP-malic enzyme 2: role of the second sphere residues in the regulatory signal transmission
title_full Structural insights into the allosteric site of Arabidopsis NADP-malic enzyme 2: role of the second sphere residues in the regulatory signal transmission
title_fullStr Structural insights into the allosteric site of Arabidopsis NADP-malic enzyme 2: role of the second sphere residues in the regulatory signal transmission
title_full_unstemmed Structural insights into the allosteric site of Arabidopsis NADP-malic enzyme 2: role of the second sphere residues in the regulatory signal transmission
title_sort structural insights into the allosteric site of arabidopsis nadp-malic enzyme 2: role of the second sphere residues in the regulatory signal transmission
publisher Springer
publishDate 2022
url http://hdl.handle.net/2133/23339
http://hdl.handle.net/2133/23339
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