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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...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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MDPI
2019
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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 |
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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 |
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