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A Novel Adenosine Kinase from Bombyx mori: Enzymatic Activity, Structure, and Biological Function.
Int J Mol Sci. 2019 Jul 31; 20(15)IJ

Abstract

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.

Authors+Show Affiliations

State Key Laboratory of Silkworm Genome Biology, Biological Science Research Center, Southwest University, Beibei, Chongqing 400715, China.State Key Laboratory of Silkworm Genome Biology, Biological Science Research Center, Southwest University, Beibei, Chongqing 400715, China.State Key Laboratory of Silkworm Genome Biology, Biological Science Research Center, Southwest University, Beibei, Chongqing 400715, China. hehuawei@swu.edu.cn. College of Biotechnology, Southwest University, Beibei, Chongqing 400715, China. hehuawei@swu.edu.cn. Chongqing Key Laboratory of Sericultural Science, Chongqing Engineering and Technology Research Center for Novel Silk Materials, Southwest University, Beibei, Chongqing 400715, China. hehuawei@swu.edu.cn.State Key Laboratory of Silkworm Genome Biology, Biological Science Research Center, Southwest University, Beibei, Chongqing 400715, China.State Key Laboratory of Silkworm Genome Biology, Biological Science Research Center, Southwest University, Beibei, Chongqing 400715, China. Chongqing Key Laboratory of Sericultural Science, Chongqing Engineering and Technology Research Center for Novel Silk Materials, Southwest University, Beibei, Chongqing 400715, China.State Key Laboratory of Silkworm Genome Biology, Biological Science Research Center, Southwest University, Beibei, Chongqing 400715, China. Chongqing Key Laboratory of Sericultural Science, Chongqing Engineering and Technology Research Center for Novel Silk Materials, Southwest University, Beibei, Chongqing 400715, China.College of Biotechnology, Southwest University, Beibei, Chongqing 400715, China. yjwang@swu.edu.cn.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31370143

Citation

Song, Kai, et al. "A Novel Adenosine Kinase From Bombyx Mori: Enzymatic Activity, Structure, and Biological Function." International Journal of Molecular Sciences, vol. 20, no. 15, 2019.
Song K, Li Y, He H, et al. A Novel Adenosine Kinase from Bombyx mori: Enzymatic Activity, Structure, and Biological Function. Int J Mol Sci. 2019;20(15).
Song, K., Li, Y., He, H., Liu, L., Zhao, P., Xia, Q., & Wang, Y. (2019). A Novel Adenosine Kinase from Bombyx mori: Enzymatic Activity, Structure, and Biological Function. International Journal of Molecular Sciences, 20(15). https://doi.org/10.3390/ijms20153732
Song K, et al. A Novel Adenosine Kinase From Bombyx Mori: Enzymatic Activity, Structure, and Biological Function. Int J Mol Sci. 2019 Jul 31;20(15) PubMed PMID: 31370143.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A Novel Adenosine Kinase from Bombyx mori: Enzymatic Activity, Structure, and Biological Function. AU - Song,Kai, AU - Li,Yu, AU - He,Huawei, AU - Liu,Lina, AU - Zhao,Ping, AU - Xia,Qingyou, AU - Wang,Yejing, Y1 - 2019/07/31/ PY - 2019/07/06/received PY - 2019/07/26/revised PY - 2019/07/29/accepted PY - 2019/8/3/entrez PY - 2019/8/3/pubmed PY - 2020/1/1/medline KW - Bombyx mori KW - adenosine kinase KW - enzymatic activity KW - structure JF - International journal of molecular sciences JO - Int J Mol Sci VL - 20 IS - 15 N2 - 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. SN - 1422-0067 UR - https://www.unboundmedicine.com/medline/citation/31370143/A_Novel_Adenosine_Kinase_from_Bombyx_mori:_Enzymatic_Activity_Structure_and_Biological_Function_ L2 - http://www.mdpi.com/resolver?pii=ijms20153732 DB - PRIME DP - Unbound Medicine ER -