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Purification, characterization, and enzyme kinetics of a glutathione S transferase from larvae of the camel tick Hyalomma dromedarii

Background Glutathione s-transferases (GSTs) perform an essential role in detoxification of xenobiotics and endogenous compounds via their conjugation to reduce glutathione. Results A GST enzyme, designated tick larvae glutathione S transferase (TLGST), was purified from larvae of the camel tick Hya...

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Published in:Journal of Genetic Engineering and Biotechnology 2023-12, Vol.21 (1), p.28-10, Article 28
Main Authors: Masoud, Hassan M. M, Helmy, Mohamed S, Darwish, Doaa A, Ibrahim, Mahmoud A
Format: Article
Language:English
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Summary:Background Glutathione s-transferases (GSTs) perform an essential role in detoxification of xenobiotics and endogenous compounds via their conjugation to reduce glutathione. Results A GST enzyme, designated tick larvae glutathione S transferase (TLGST), was purified from larvae of the camel tick Hyalomma dromedarii via ammonium sulfate precipitation, glutathione-Sepharose affinity column and Sephacryl S-300 chromatography. TLGST-specific activity was found to be 1.56 Umg.sup.-1 which represents 39 folds and 32.2% recovery. The molecular weight of TLGST purified from camel tick larvae was found as 42 kDa by gel filtration. TLGST has a pI value of 6.9 and was found a heterodimeric protein of 28 and 14 kDa subunits as detected on SDS-PAGE. The Lineweaver-Burk plot calculated the km for CDNB to be 0.43 mM with Vmax value of 9.2 Umg.sup.-1. TLGST exhibited its optimal activity at pH 7.9. Co.sup.2+, Ni.sup.2+ and Mn.sup.2+ increased the activity of TLGST while Ca.sup.2+, Cu.sup.2+, Fe.sup.2+ and Zn.sup.2+ inhibited it. TLGST was inhibited by cumene hydroperoxide, p-hydroxymercuribenzoate, lithocholic acid, hematin, triphenyltin chloride, p-chloromercuribenzoic acid (pCMB), N-p-Tosyl-L-phenylalanine chloromethyl ketone (TPCK), iodoacetamide, EDTA and quercetin. pCMB inhibited TLGST competitively with Ki value of 0.3 mM. Conclusions These findings will help to understand the various physiologic conditions of ticks and targeting TLGST could be significant tool for development of prospective vaccines against ticks as a bio-control strategy to overcome the rapid grows in pesticide-resistant tick populations. Graphical
ISSN:1687-157X
2090-5920
DOI:10.1186/s43141-023-00486-w