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Heterologous overexpression of active hexokinases from microsporidia Nosema bombycis and Nosema ceranae confirms their ability to phosphorylate host glucose.

التفاصيل البيبلوغرافية
العنوان: Heterologous overexpression of active hexokinases from microsporidia Nosema bombycis and Nosema ceranae confirms their ability to phosphorylate host glucose.
المؤلفون: Dolgikh VV; Laboratory of Microbiological Control, All-Russian Institute for Plant Protection, Podbelskogo shosse, 3, Pushkin, St. Petersburg, 196608, Russia. dol1slav@yahoo.com., Tsarev AA; Laboratory of Microbiological Control, All-Russian Institute for Plant Protection, Podbelskogo shosse, 3, Pushkin, St. Petersburg, 196608, Russia., Timofeev SA; Laboratory of Microbiological Control, All-Russian Institute for Plant Protection, Podbelskogo shosse, 3, Pushkin, St. Petersburg, 196608, Russia., Zhuravlyov VS; Laboratory of Microbiological Control, All-Russian Institute for Plant Protection, Podbelskogo shosse, 3, Pushkin, St. Petersburg, 196608, Russia.
المصدر: Parasitology research [Parasitol Res] 2019 May; Vol. 118 (5), pp. 1511-1518. Date of Electronic Publication: 2019 Mar 13.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: Springer International Country of Publication: Germany NLM ID: 8703571 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1432-1955 (Electronic) Linking ISSN: 09320113 NLM ISO Abbreviation: Parasitol Res Subsets: MEDLINE
أسماء مطبوعة: Original Publication: Berlin : Springer International, c1987-
مواضيع طبية MeSH: Bees/*parasitology , Bombyx/*parasitology , Hexokinase/*biosynthesis , Nosema/*metabolism, Animals ; Escherichia coli/genetics ; Glucose/metabolism ; Hexokinase/genetics ; Nosema/classification ; Nosema/isolation & purification ; Phosphorylation
مستخلص: The secretion of hexokinases (HKs) by microsporidia followed by their accumulation in insect host nuclei suggests that these enzymes play regulatory and catalytic roles in infected cells. To confirm whether HKs exert catalytic functions in insect cells, we expressed in E. coli the functionally active HKs of two entomopathogenic microsporidia, Nosema bombycis and Nosema ceranae, that cause silkworm and honey bee nosematoses. N. bombycis HK with C-terminal polyHis tag and N. ceranae enzyme with N-terminal polyHis tag were cloned into pOPE101 and pRSET vectors, respectively, and overexpressed. Specific activities of N. bombycis and N. ceranae enzymes isolated by metal chelate affinity chromatography were 29.2 ± 0.5 and 60.2 ± 1.2 U/mg protein at an optimal pH range of 8.5-9.5. The kinetic characteristics of the recombinant enzymes were similar to those of HKs from other parasitic and free-living organisms. N. bombycis HK demonstrated Km 0.07 ± 0.01 mM and k cat 1726 min -1 for glucose, and Km 0.39 ± 0.05 mM and k cat 1976 min -1 for ATP, at pH 8.8. N. ceranae HK showed Km 0.3 ± 0.04 mM and k cat 3293 min -1 for glucose, and Km 1.15 ± 0.11 mM and k cat 3732 min -1 for ATP, at the same pH value. These data demonstrate the capability of microsporidia-secreted HKs to phosphorylate glucose in infected cells, suggesting that they actively mediate the effects of the parasite on host metabolism. The present findings justify further study of the enzymes as targets to suppress the intracellular development of silkworm and honey bee pathogens.
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فهرسة مساهمة: Keywords: Enzyme assay; Heterologous expression; Hexokinase; Microsporidia; Nosema bombycis; Nosema ceranae
المشرفين على المادة: EC 2.7.1.1 (Hexokinase)
IY9XDZ35W2 (Glucose)
تواريخ الأحداث: Date Created: 20190314 Date Completed: 20190618 Latest Revision: 20200225
رمز التحديث: 20231215
DOI: 10.1007/s00436-019-06279-w
PMID: 30863897
قاعدة البيانات: MEDLINE
الوصف
تدمد:1432-1955
DOI:10.1007/s00436-019-06279-w