Identification and characterization of acid and alkaline phosphatases and protein phosphatases in L. catesbeianus tail during metamorphosis
Autor(a) principal: | |
---|---|
Data de Publicação: | 2021 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1007/s11756-021-00877-9 http://hdl.handle.net/11449/229491 |
Resumo: | Amphibian metamorphosis is a tightly regulated transformation involving the participation of hormones and other biomolecules in cell death and tail absorption. Among the regulators, phosphate is essential for various processes, such as cell death and survival and enzyme activity regulation. Therefore, identification and characterization of phosphatases in L. catesbeianus tail may contribute to understand the events involved in cell death and nutrient release during metamorphosis. Differential centrifugation was used to separate soluble proteins from membrane proteins and analyzed by phosphomonohydrolases, serine/threonine protein phosphatase 2A (PP2A), and protein tyrosine phosphatase (PTP) assays. Mitochondrial fractioning was used to evaluate PP2A and alkaline phosphatase activities. Tandem mass spectrometry (MS/MS) analysis was performed using the crude extract. Phosphomonohydrolase activity was assayed by p-nitrophenylphosphate (pNPP) hydrolysis, whereas PP2A and PTP were assayed by peptides phosphorylated in threonine and tyrosine, respectively; inhibitor-2 was used to identify the serine/threonine protein phosphatase type 1 (PP1). The enzymatic activities and kinetic parameters of pNPP hydrolysis revealed three distinct phosphomonohydrolases. MS/MS analysis of the crude extract revealed three protein phosphatases, viz., PP2A, PP1, and a PTP, which was confirmed by in vitro assays. The results may relate PP2A activity to membrane-bound alkaline phosphatase, PTP activity to soluble acid phosphatase, and PP1 activity to membrane acid phosphatase, although more detailed studies are needed to confirm this hypothesis. We propose a model providing information on the role of PP1 and PP2A in the signaling events leading to cell death and the role of these enzymes in anuran tail absorption during metamorphosis. |
id |
UNSP_694439275b2094193444b0b7e61dda8a |
---|---|
oai_identifier_str |
oai:repositorio.unesp.br:11449/229491 |
network_acronym_str |
UNSP |
network_name_str |
Repositório Institucional da UNESP |
repository_id_str |
2946 |
spelling |
Identification and characterization of acid and alkaline phosphatases and protein phosphatases in L. catesbeianus tail during metamorphosisAnuraCell deathMetamorphosisMitochondriaPhosphatasesPP1PP2AAmphibian metamorphosis is a tightly regulated transformation involving the participation of hormones and other biomolecules in cell death and tail absorption. Among the regulators, phosphate is essential for various processes, such as cell death and survival and enzyme activity regulation. Therefore, identification and characterization of phosphatases in L. catesbeianus tail may contribute to understand the events involved in cell death and nutrient release during metamorphosis. Differential centrifugation was used to separate soluble proteins from membrane proteins and analyzed by phosphomonohydrolases, serine/threonine protein phosphatase 2A (PP2A), and protein tyrosine phosphatase (PTP) assays. Mitochondrial fractioning was used to evaluate PP2A and alkaline phosphatase activities. Tandem mass spectrometry (MS/MS) analysis was performed using the crude extract. Phosphomonohydrolase activity was assayed by p-nitrophenylphosphate (pNPP) hydrolysis, whereas PP2A and PTP were assayed by peptides phosphorylated in threonine and tyrosine, respectively; inhibitor-2 was used to identify the serine/threonine protein phosphatase type 1 (PP1). The enzymatic activities and kinetic parameters of pNPP hydrolysis revealed three distinct phosphomonohydrolases. MS/MS analysis of the crude extract revealed three protein phosphatases, viz., PP2A, PP1, and a PTP, which was confirmed by in vitro assays. The results may relate PP2A activity to membrane-bound alkaline phosphatase, PTP activity to soluble acid phosphatase, and PP1 activity to membrane acid phosphatase, although more detailed studies are needed to confirm this hypothesis. We propose a model providing information on the role of PP1 and PP2A in the signaling events leading to cell death and the role of these enzymes in anuran tail absorption during metamorphosis.Department of Biological and Health Sciences University of Araraquara (UNIARA)Chemistry Institute Department of Biochemistry and Organic Chemistry São Paulo State University (UNESP)School of Agricultural and Veterinarian Sciences Department of Technology São Paulo State University (UNESP)Instituto Municipal de Ensino Superior de Bebedouro “Victório Cardassi” (IMESB), Rua Nelson Domingos Madeira, 300Chemistry Institute Department of Biochemistry and Organic Chemistry São Paulo State University (UNESP)School of Agricultural and Veterinarian Sciences Department of Technology São Paulo State University (UNESP)University of Araraquara (UNIARA)Universidade Estadual Paulista (UNESP)Instituto Municipal de Ensino Superior de Bebedouro “Victório Cardassi” (IMESB)Gonçalves, Adriano Marques [UNESP]Santana, Caroline Carla [UNESP]Santos, Luiz Flávio José Dos [UNESP]Colosio, Rafael Rodrigues [UNESP]Balbuena, Tiago Santana [UNESP]Pizauro, João Martins [UNESP]2022-04-29T08:32:46Z2022-04-29T08:32:46Z2021-01-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://dx.doi.org/10.1007/s11756-021-00877-9Biologia.1336-95630006-3088http://hdl.handle.net/11449/22949110.1007/s11756-021-00877-92-s2.0-85114615264Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengBiologiainfo:eu-repo/semantics/openAccess2022-04-29T08:32:47Zoai:repositorio.unesp.br:11449/229491Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T19:12:27.417139Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Identification and characterization of acid and alkaline phosphatases and protein phosphatases in L. catesbeianus tail during metamorphosis |
title |
Identification and characterization of acid and alkaline phosphatases and protein phosphatases in L. catesbeianus tail during metamorphosis |
spellingShingle |
Identification and characterization of acid and alkaline phosphatases and protein phosphatases in L. catesbeianus tail during metamorphosis Gonçalves, Adriano Marques [UNESP] Anura Cell death Metamorphosis Mitochondria Phosphatases PP1 PP2A |
title_short |
Identification and characterization of acid and alkaline phosphatases and protein phosphatases in L. catesbeianus tail during metamorphosis |
title_full |
Identification and characterization of acid and alkaline phosphatases and protein phosphatases in L. catesbeianus tail during metamorphosis |
title_fullStr |
Identification and characterization of acid and alkaline phosphatases and protein phosphatases in L. catesbeianus tail during metamorphosis |
title_full_unstemmed |
Identification and characterization of acid and alkaline phosphatases and protein phosphatases in L. catesbeianus tail during metamorphosis |
title_sort |
Identification and characterization of acid and alkaline phosphatases and protein phosphatases in L. catesbeianus tail during metamorphosis |
author |
Gonçalves, Adriano Marques [UNESP] |
author_facet |
Gonçalves, Adriano Marques [UNESP] Santana, Caroline Carla [UNESP] Santos, Luiz Flávio José Dos [UNESP] Colosio, Rafael Rodrigues [UNESP] Balbuena, Tiago Santana [UNESP] Pizauro, João Martins [UNESP] |
author_role |
author |
author2 |
Santana, Caroline Carla [UNESP] Santos, Luiz Flávio José Dos [UNESP] Colosio, Rafael Rodrigues [UNESP] Balbuena, Tiago Santana [UNESP] Pizauro, João Martins [UNESP] |
author2_role |
author author author author author |
dc.contributor.none.fl_str_mv |
University of Araraquara (UNIARA) Universidade Estadual Paulista (UNESP) Instituto Municipal de Ensino Superior de Bebedouro “Victório Cardassi” (IMESB) |
dc.contributor.author.fl_str_mv |
Gonçalves, Adriano Marques [UNESP] Santana, Caroline Carla [UNESP] Santos, Luiz Flávio José Dos [UNESP] Colosio, Rafael Rodrigues [UNESP] Balbuena, Tiago Santana [UNESP] Pizauro, João Martins [UNESP] |
dc.subject.por.fl_str_mv |
Anura Cell death Metamorphosis Mitochondria Phosphatases PP1 PP2A |
topic |
Anura Cell death Metamorphosis Mitochondria Phosphatases PP1 PP2A |
description |
Amphibian metamorphosis is a tightly regulated transformation involving the participation of hormones and other biomolecules in cell death and tail absorption. Among the regulators, phosphate is essential for various processes, such as cell death and survival and enzyme activity regulation. Therefore, identification and characterization of phosphatases in L. catesbeianus tail may contribute to understand the events involved in cell death and nutrient release during metamorphosis. Differential centrifugation was used to separate soluble proteins from membrane proteins and analyzed by phosphomonohydrolases, serine/threonine protein phosphatase 2A (PP2A), and protein tyrosine phosphatase (PTP) assays. Mitochondrial fractioning was used to evaluate PP2A and alkaline phosphatase activities. Tandem mass spectrometry (MS/MS) analysis was performed using the crude extract. Phosphomonohydrolase activity was assayed by p-nitrophenylphosphate (pNPP) hydrolysis, whereas PP2A and PTP were assayed by peptides phosphorylated in threonine and tyrosine, respectively; inhibitor-2 was used to identify the serine/threonine protein phosphatase type 1 (PP1). The enzymatic activities and kinetic parameters of pNPP hydrolysis revealed three distinct phosphomonohydrolases. MS/MS analysis of the crude extract revealed three protein phosphatases, viz., PP2A, PP1, and a PTP, which was confirmed by in vitro assays. The results may relate PP2A activity to membrane-bound alkaline phosphatase, PTP activity to soluble acid phosphatase, and PP1 activity to membrane acid phosphatase, although more detailed studies are needed to confirm this hypothesis. We propose a model providing information on the role of PP1 and PP2A in the signaling events leading to cell death and the role of these enzymes in anuran tail absorption during metamorphosis. |
publishDate |
2021 |
dc.date.none.fl_str_mv |
2021-01-01 2022-04-29T08:32:46Z 2022-04-29T08:32:46Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1007/s11756-021-00877-9 Biologia. 1336-9563 0006-3088 http://hdl.handle.net/11449/229491 10.1007/s11756-021-00877-9 2-s2.0-85114615264 |
url |
http://dx.doi.org/10.1007/s11756-021-00877-9 http://hdl.handle.net/11449/229491 |
identifier_str_mv |
Biologia. 1336-9563 0006-3088 10.1007/s11756-021-00877-9 2-s2.0-85114615264 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Biologia |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1808129032938586112 |