Cargando…

A Novel Adenosine Kinase from Bombyx mori: Enzymatic Activity, Structure, and Biological Function

Adenosine kinase (ADK) is the first enzyme in the adenosine remediation pathway that catalyzes adenosine phosphorylation into adenosine monophosphate, thus regulating adenosine homeostasis in cells. To obtain new insights into ADK from Bombyx mori (BmADK), we obtained recombinant BmADK, and analyzed...

Descripción completa

Detalles Bibliográficos
Autores principales: Song, Kai, Li, Yu, He, Huawei, Liu, Lina, Zhao, Ping, Xia, Qingyou, Wang, Yejing
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6695918/
https://www.ncbi.nlm.nih.gov/pubmed/31370143
http://dx.doi.org/10.3390/ijms20153732
_version_ 1783444147787530240
author Song, Kai
Li, Yu
He, Huawei
Liu, Lina
Zhao, Ping
Xia, Qingyou
Wang, Yejing
author_facet Song, Kai
Li, Yu
He, Huawei
Liu, Lina
Zhao, Ping
Xia, Qingyou
Wang, Yejing
author_sort Song, Kai
collection PubMed
description Adenosine kinase (ADK) is the first enzyme in the adenosine remediation pathway that catalyzes adenosine phosphorylation into adenosine monophosphate, thus regulating adenosine homeostasis in cells. To obtain new insights into ADK from Bombyx mori (BmADK), we obtained recombinant BmADK, and analyzed its activity, structure, and function. Gel-filtration showed BmADK was a monomer with molecular weight of approximately 38 kDa. Circular dichroism spectra indicated BmADK had 36.8% α-helix and 29.9% β-strand structures, respectively. The structure of BmADK was stable in pH 5.0–11.0, and not affected under 30 °C. The melting temperature and the enthalpy and entropy changes in the thermal transition of BmADK were 46.51 ± 0.50 °C, 253.43 ± 0.20 KJ/mol, and 0.79 ± 0.01 KJ/(mol·K), respectively. Site-directed mutagenesis demonstrated G68, S201, E229, and D303 were key amino acids for BmADK structure and activity. In particular, S201A mutation significantly increased the α-helix content of BmADK and its activity. BmADK was located in the cytoplasm and highly expressed in the silk gland during the pre-pupal stage. RNA interference revealed the downregulation of BmADK decreased ATG-8, Caspase-9, Ec-R, E74A, and Br-C expression, indicating it was likely involved in 20E signaling, apoptosis, and autophagy to regulate silk gland degeneration and silkworm metamorphosis. Our study greatly expanded the knowledge on the activity, structure, and role of ADK.
format Online
Article
Text
id pubmed-6695918
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-66959182019-09-05 A Novel Adenosine Kinase from Bombyx mori: Enzymatic Activity, Structure, and Biological Function Song, Kai Li, Yu He, Huawei Liu, Lina Zhao, Ping Xia, Qingyou Wang, Yejing Int J Mol Sci Article Adenosine kinase (ADK) is the first enzyme in the adenosine remediation pathway that catalyzes adenosine phosphorylation into adenosine monophosphate, thus regulating adenosine homeostasis in cells. To obtain new insights into ADK from Bombyx mori (BmADK), we obtained recombinant BmADK, and analyzed its activity, structure, and function. Gel-filtration showed BmADK was a monomer with molecular weight of approximately 38 kDa. Circular dichroism spectra indicated BmADK had 36.8% α-helix and 29.9% β-strand structures, respectively. The structure of BmADK was stable in pH 5.0–11.0, and not affected under 30 °C. The melting temperature and the enthalpy and entropy changes in the thermal transition of BmADK were 46.51 ± 0.50 °C, 253.43 ± 0.20 KJ/mol, and 0.79 ± 0.01 KJ/(mol·K), respectively. Site-directed mutagenesis demonstrated G68, S201, E229, and D303 were key amino acids for BmADK structure and activity. In particular, S201A mutation significantly increased the α-helix content of BmADK and its activity. BmADK was located in the cytoplasm and highly expressed in the silk gland during the pre-pupal stage. RNA interference revealed the downregulation of BmADK decreased ATG-8, Caspase-9, Ec-R, E74A, and Br-C expression, indicating it was likely involved in 20E signaling, apoptosis, and autophagy to regulate silk gland degeneration and silkworm metamorphosis. Our study greatly expanded the knowledge on the activity, structure, and role of ADK. MDPI 2019-07-31 /pmc/articles/PMC6695918/ /pubmed/31370143 http://dx.doi.org/10.3390/ijms20153732 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Song, Kai
Li, Yu
He, Huawei
Liu, Lina
Zhao, Ping
Xia, Qingyou
Wang, Yejing
A Novel Adenosine Kinase from Bombyx mori: Enzymatic Activity, Structure, and Biological Function
title A Novel Adenosine Kinase from Bombyx mori: Enzymatic Activity, Structure, and Biological Function
title_full A Novel Adenosine Kinase from Bombyx mori: Enzymatic Activity, Structure, and Biological Function
title_fullStr A Novel Adenosine Kinase from Bombyx mori: Enzymatic Activity, Structure, and Biological Function
title_full_unstemmed A Novel Adenosine Kinase from Bombyx mori: Enzymatic Activity, Structure, and Biological Function
title_short A Novel Adenosine Kinase from Bombyx mori: Enzymatic Activity, Structure, and Biological Function
title_sort novel adenosine kinase from bombyx mori: enzymatic activity, structure, and biological function
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6695918/
https://www.ncbi.nlm.nih.gov/pubmed/31370143
http://dx.doi.org/10.3390/ijms20153732
work_keys_str_mv AT songkai anoveladenosinekinasefrombombyxmorienzymaticactivitystructureandbiologicalfunction
AT liyu anoveladenosinekinasefrombombyxmorienzymaticactivitystructureandbiologicalfunction
AT hehuawei anoveladenosinekinasefrombombyxmorienzymaticactivitystructureandbiologicalfunction
AT liulina anoveladenosinekinasefrombombyxmorienzymaticactivitystructureandbiologicalfunction
AT zhaoping anoveladenosinekinasefrombombyxmorienzymaticactivitystructureandbiologicalfunction
AT xiaqingyou anoveladenosinekinasefrombombyxmorienzymaticactivitystructureandbiologicalfunction
AT wangyejing anoveladenosinekinasefrombombyxmorienzymaticactivitystructureandbiologicalfunction
AT songkai noveladenosinekinasefrombombyxmorienzymaticactivitystructureandbiologicalfunction
AT liyu noveladenosinekinasefrombombyxmorienzymaticactivitystructureandbiologicalfunction
AT hehuawei noveladenosinekinasefrombombyxmorienzymaticactivitystructureandbiologicalfunction
AT liulina noveladenosinekinasefrombombyxmorienzymaticactivitystructureandbiologicalfunction
AT zhaoping noveladenosinekinasefrombombyxmorienzymaticactivitystructureandbiologicalfunction
AT xiaqingyou noveladenosinekinasefrombombyxmorienzymaticactivitystructureandbiologicalfunction
AT wangyejing noveladenosinekinasefrombombyxmorienzymaticactivitystructureandbiologicalfunction