Biochemical and structural characterization of a novel arginine kinase from the spider Polybetes pythagoricus

Autores
Laino, Aldana; Lopez Zavala, Alonso A.; Garcia, Carlos Fernando; Carrasco Miranda, Jesus S.; Santana, Marianela; Stojanoff, Vivian; Sotelo Mundo, Rogerio Rafael; Garcia, Carlos Fernando
Año de publicación
2017
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Energy buffering systems are key for homeostasis during variations in energy supply. Spiders are the most important predators for insects and therefore key in terrestrial ecosystems. From biomedical interest, spiders are important for their venoms and as a source of potent allergens, such as arginine kinase (AK, EC 2.7.3.3). AK is an enzyme crucial for energy metabolism, keeping the pool of phosphagens in invertebrates, and also an allergen for humans. In this work, we studied AK from the Argentininan spider Polybetes pythagoricus (PpAK), from its complementary DNA to the crystal structure. The PpAK cDNA from muscle was cloned, and it is comprised of 1068 nucleotides that encode a 384-amino acids protein, similar to other invertebrate AKs. The apparent Michaelis-Menten kinetic constant (Km) was 1.7 mM with a kcat of 75 s-1. Two crystal structures are presented, the apoPvAK and PpAK bound to arginine, both in the open conformation with the active site lid (residues 310-320) completely disordered. The guanidino group binding site in the apo structure appears to be organized to accept the arginine substrate. Finally, these results contribute to knowledge of mechanistic details of the function of arginine kinase.
Fil: Laino, Aldana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; Argentina
Fil: Lopez Zavala, Alonso A.. Universidad de Sonora; México
Fil: Garcia, Carlos Fernando. Centro de Investigación en Alimentación y Desarrollo; México
Fil: Carrasco Miranda, Jesus S.. Centro de Investigación en Alimentación y Desarrollo; México
Fil: Santana, Marianela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; Argentina
Fil: Stojanoff, Vivian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; Argentina
Fil: Sotelo Mundo, Rogerio Rafael. Centro de Investigación en Alimentación y Desarrollo; México. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Garcia, Carlos Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; Argentina
Materia
ALLERGEN
ARACHNIDA
ARGININE KINASE
ARTHROPODA
CDNA CLONING
CRYSTAL STRUCTURE
OPEN CONFORMATION
PHOSPHAGEN
POLYBETES PYTAGORICUS
SPIDER
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/51679
Acceder
id CONICETDig_3ebea480a1e9fd442e3b53b0f007ee1e
oai_identifier_str oai:ri.conicet.gov.ar:11336/51679
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Biochemical and structural characterization of a novel arginine kinase from the spider Polybetes pythagoricusLaino, AldanaLopez Zavala, Alonso A.Garcia, Carlos FernandoCarrasco Miranda, Jesus S.Santana, MarianelaStojanoff, VivianSotelo Mundo, Rogerio RafaelGarcia, Carlos FernandoALLERGENARACHNIDAARGININE KINASEARTHROPODACDNA CLONINGCRYSTAL STRUCTUREOPEN CONFORMATIONPHOSPHAGENPOLYBETES PYTAGORICUSSPIDERhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Energy buffering systems are key for homeostasis during variations in energy supply. Spiders are the most important predators for insects and therefore key in terrestrial ecosystems. From biomedical interest, spiders are important for their venoms and as a source of potent allergens, such as arginine kinase (AK, EC 2.7.3.3). AK is an enzyme crucial for energy metabolism, keeping the pool of phosphagens in invertebrates, and also an allergen for humans. In this work, we studied AK from the Argentininan spider Polybetes pythagoricus (PpAK), from its complementary DNA to the crystal structure. The PpAK cDNA from muscle was cloned, and it is comprised of 1068 nucleotides that encode a 384-amino acids protein, similar to other invertebrate AKs. The apparent Michaelis-Menten kinetic constant (Km) was 1.7 mM with a kcat of 75 s-1. Two crystal structures are presented, the apoPvAK and PpAK bound to arginine, both in the open conformation with the active site lid (residues 310-320) completely disordered. The guanidino group binding site in the apo structure appears to be organized to accept the arginine substrate. Finally, these results contribute to knowledge of mechanistic details of the function of arginine kinase.Fil: Laino, Aldana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; ArgentinaFil: Lopez Zavala, Alonso A.. Universidad de Sonora; MéxicoFil: Garcia, Carlos Fernando. Centro de Investigación en Alimentación y Desarrollo; MéxicoFil: Carrasco Miranda, Jesus S.. Centro de Investigación en Alimentación y Desarrollo; MéxicoFil: Santana, Marianela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; ArgentinaFil: Stojanoff, Vivian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; ArgentinaFil: Sotelo Mundo, Rogerio Rafael. Centro de Investigación en Alimentación y Desarrollo; México. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Garcia, Carlos Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; ArgentinaPeerJ Inc.2017-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/51679Laino, Aldana; Lopez Zavala, Alonso A.; Garcia, Carlos Fernando; Carrasco Miranda, Jesus S.; Santana, Marianela; et al.; Biochemical and structural characterization of a novel arginine kinase from the spider Polybetes pythagoricus; PeerJ Inc.; PeerJ; 2017; 9; 12-2017; 1-192167-8359CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.7717/peerj.3787info:eu-repo/semantics/altIdentifier/url/https://peerj.com/articles/3787/info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-10T13:08:32Zoai:ri.conicet.gov.ar:11336/51679instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-10 13:08:32.904CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Biochemical and structural characterization of a novel arginine kinase from the spider Polybetes pythagoricus
title Biochemical and structural characterization of a novel arginine kinase from the spider Polybetes pythagoricus
spellingShingle Biochemical and structural characterization of a novel arginine kinase from the spider Polybetes pythagoricus
Laino, Aldana
ALLERGEN
ARACHNIDA
ARGININE KINASE
ARTHROPODA
CDNA CLONING
CRYSTAL STRUCTURE
OPEN CONFORMATION
PHOSPHAGEN
POLYBETES PYTAGORICUS
SPIDER
title_short Biochemical and structural characterization of a novel arginine kinase from the spider Polybetes pythagoricus
title_full Biochemical and structural characterization of a novel arginine kinase from the spider Polybetes pythagoricus
title_fullStr Biochemical and structural characterization of a novel arginine kinase from the spider Polybetes pythagoricus
title_full_unstemmed Biochemical and structural characterization of a novel arginine kinase from the spider Polybetes pythagoricus
title_sort Biochemical and structural characterization of a novel arginine kinase from the spider Polybetes pythagoricus
dc.creator.none.fl_str_mv Laino, Aldana
Lopez Zavala, Alonso A.
Garcia, Carlos Fernando
Carrasco Miranda, Jesus S.
Santana, Marianela
Stojanoff, Vivian
Sotelo Mundo, Rogerio Rafael
Garcia, Carlos Fernando
author Laino, Aldana
author_facet Laino, Aldana
Lopez Zavala, Alonso A.
Garcia, Carlos Fernando
Carrasco Miranda, Jesus S.
Santana, Marianela
Stojanoff, Vivian
Sotelo Mundo, Rogerio Rafael
author_role author
author2 Lopez Zavala, Alonso A.
Garcia, Carlos Fernando
Carrasco Miranda, Jesus S.
Santana, Marianela
Stojanoff, Vivian
Sotelo Mundo, Rogerio Rafael
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv ALLERGEN
ARACHNIDA
ARGININE KINASE
ARTHROPODA
CDNA CLONING
CRYSTAL STRUCTURE
OPEN CONFORMATION
PHOSPHAGEN
POLYBETES PYTAGORICUS
SPIDER
topic ALLERGEN
ARACHNIDA
ARGININE KINASE
ARTHROPODA
CDNA CLONING
CRYSTAL STRUCTURE
OPEN CONFORMATION
PHOSPHAGEN
POLYBETES PYTAGORICUS
SPIDER
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Energy buffering systems are key for homeostasis during variations in energy supply. Spiders are the most important predators for insects and therefore key in terrestrial ecosystems. From biomedical interest, spiders are important for their venoms and as a source of potent allergens, such as arginine kinase (AK, EC 2.7.3.3). AK is an enzyme crucial for energy metabolism, keeping the pool of phosphagens in invertebrates, and also an allergen for humans. In this work, we studied AK from the Argentininan spider Polybetes pythagoricus (PpAK), from its complementary DNA to the crystal structure. The PpAK cDNA from muscle was cloned, and it is comprised of 1068 nucleotides that encode a 384-amino acids protein, similar to other invertebrate AKs. The apparent Michaelis-Menten kinetic constant (Km) was 1.7 mM with a kcat of 75 s-1. Two crystal structures are presented, the apoPvAK and PpAK bound to arginine, both in the open conformation with the active site lid (residues 310-320) completely disordered. The guanidino group binding site in the apo structure appears to be organized to accept the arginine substrate. Finally, these results contribute to knowledge of mechanistic details of the function of arginine kinase.
Fil: Laino, Aldana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; Argentina
Fil: Lopez Zavala, Alonso A.. Universidad de Sonora; México
Fil: Garcia, Carlos Fernando. Centro de Investigación en Alimentación y Desarrollo; México
Fil: Carrasco Miranda, Jesus S.. Centro de Investigación en Alimentación y Desarrollo; México
Fil: Santana, Marianela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; Argentina
Fil: Stojanoff, Vivian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; Argentina
Fil: Sotelo Mundo, Rogerio Rafael. Centro de Investigación en Alimentación y Desarrollo; México. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Garcia, Carlos Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; Argentina
description Energy buffering systems are key for homeostasis during variations in energy supply. Spiders are the most important predators for insects and therefore key in terrestrial ecosystems. From biomedical interest, spiders are important for their venoms and as a source of potent allergens, such as arginine kinase (AK, EC 2.7.3.3). AK is an enzyme crucial for energy metabolism, keeping the pool of phosphagens in invertebrates, and also an allergen for humans. In this work, we studied AK from the Argentininan spider Polybetes pythagoricus (PpAK), from its complementary DNA to the crystal structure. The PpAK cDNA from muscle was cloned, and it is comprised of 1068 nucleotides that encode a 384-amino acids protein, similar to other invertebrate AKs. The apparent Michaelis-Menten kinetic constant (Km) was 1.7 mM with a kcat of 75 s-1. Two crystal structures are presented, the apoPvAK and PpAK bound to arginine, both in the open conformation with the active site lid (residues 310-320) completely disordered. The guanidino group binding site in the apo structure appears to be organized to accept the arginine substrate. Finally, these results contribute to knowledge of mechanistic details of the function of arginine kinase.
publishDate 2017
dc.date.none.fl_str_mv 2017-12
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/51679
Laino, Aldana; Lopez Zavala, Alonso A.; Garcia, Carlos Fernando; Carrasco Miranda, Jesus S.; Santana, Marianela; et al.; Biochemical and structural characterization of a novel arginine kinase from the spider Polybetes pythagoricus; PeerJ Inc.; PeerJ; 2017; 9; 12-2017; 1-19
2167-8359
CONICET Digital
CONICET
url http://hdl.handle.net/11336/51679
identifier_str_mv Laino, Aldana; Lopez Zavala, Alonso A.; Garcia, Carlos Fernando; Carrasco Miranda, Jesus S.; Santana, Marianela; et al.; Biochemical and structural characterization of a novel arginine kinase from the spider Polybetes pythagoricus; PeerJ Inc.; PeerJ; 2017; 9; 12-2017; 1-19
2167-8359
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.7717/peerj.3787
info:eu-repo/semantics/altIdentifier/url/https://peerj.com/articles/3787/
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv PeerJ Inc.
publisher.none.fl_str_mv PeerJ Inc.
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 12.993085

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