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

Autor: Masoud HMM; Molecular Biology Department, National Research Centre, El-Tahrir St, Dokki, Giza, Egypt. hssnmasoud@yahoo.com.; Proteome Research Laboratory, Central Laboratories Network and Centers of Excellence, National Research Centre, El-Tahrir St, Dokki, Giza, Egypt. hssnmasoud@yahoo.com., Helmy MS; Molecular Biology Department, National Research Centre, El-Tahrir St, Dokki, Giza, Egypt.; Proteome Research Laboratory, Central Laboratories Network and Centers of Excellence, National Research Centre, El-Tahrir St, Dokki, Giza, Egypt., Darwish DA; Molecular Biology Department, National Research Centre, El-Tahrir St, Dokki, Giza, Egypt.; Proteome Research Laboratory, Central Laboratories Network and Centers of Excellence, National Research Centre, El-Tahrir St, Dokki, Giza, Egypt., Ibrahim MA; Molecular Biology Department, National Research Centre, El-Tahrir St, Dokki, Giza, Egypt.; Proteome Research Laboratory, Central Laboratories Network and Centers of Excellence, National Research Centre, El-Tahrir St, Dokki, Giza, Egypt.
Jazyk: angličtina
Zdroj: Journal, genetic engineering & biotechnology [J Genet Eng Biotechnol] 2023 Mar 08; Vol. 21 (1), pp. 28. Date of Electronic Publication: 2023 Mar 08.
DOI: 10.1186/s43141-023-00486-w
Abstrakt: 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 -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 -1 . TLGST exhibited its optimal activity at pH 7.9. Co 2+ , Ni 2+ and Mn 2+ increased the activity of TLGST while Ca 2+ , Cu 2+ , Fe 2+ and Zn 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.
(© 2023. The Author(s).)
Databáze: MEDLINE
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