Interaction of nanoparticles with arginine kinase from Trypanosomabrucei: Kinetic and mechanistic evaluation

Adeyemi, O. S. and Whiteley, C.G. (2013) Interaction of nanoparticles with arginine kinase from Trypanosomabrucei: Kinetic and mechanistic evaluation. InternationalJournalofBiologicalMacromolecules, 62. pp. 450-456. ISSN 0141-8130/$

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Abstract

Argininekinaseisnotonlyabsentfrommammalianhostsbutiscriticaltothesurvivaloftrypanosomesunderstressfulconditionsandconsequentlyitsinhibitionmayleadtoaneffectivetreatmentfortry-panosomiasis.TheHis-taggedenzymewasclonedfromTrypanosomabruceigenomicDNA,expressedinEscherichiacoliBL21DE3cellsandpurifiedonaNi-affinitycolumnandbyFPLConaSuperdex200HR.Theenzymehadaspecificactivityof2.92 molmin−1mgprotein−1,molecularmassof40kDa,temper-atureandpHoptimaof30◦Cand7.8,andKmandVmaxas2.94mMand0.161�molml−1min−1(argininesubstrate).Theinteractionoftheenzymewithsilverandgoldnanoparticlesshowedanon-competitiveinhibitionwith,respectively,75%and62%decreaseinactivity;Kivaluesrangedfrom1.5nM(Ag)to3.1nM(Au).AmechanismforthisinhibitionwasbyinteractionwithCys271positioned3.3 ̊AfromthereactiveNH1ofsubstratearginine.ThiscysteinecontrolselectrophilicandnucleophiliccharacteroftheguanidiniumgroupthatiscrucialforenzymaticphosphoryltransferbetweenADPandATP.

Item Type:	Article
Subjects:	Q Science > QH Natural history > QH426 Genetics
Depositing User:	Mr DIGITAL CONTENT CREATOR LMU
Date Deposited:	17 Sep 2019 09:24
Last Modified:	17 Sep 2019 09:24
URI:	https://eprints.lmu.edu.ng/id/eprint/2228

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