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Title 

Towards binding mechanism of Cu2+ on creatine kinase from pelodiscus sinensis : molecular dynamics simulation integrating inhibition kinetics study

Authors 

Y CaiJinhyuk LeeW WangY D ParkG Y Qian

Publisher 

Bentham Science Publishers

Issue Date 

2017

Citation 

Protein and Peptide Letters

Keywords 

Creatine kinaseCu2+InactivationMD simulationOsmolytesPelodiscus sinensis

Abstract 

Background: Cu2+ is well known to play important roles in living organisms having bifacial distinction: essential microelement that is necessary for a wide range of metabolic processes but hyper-accumulation of Cu2+ can be toxic. The physiological function of Cu2+ in ectothermic animals such as Pelodiscus sinensis (Chinese soft-shelled turtle) has not been elucidated. Objective: In this study, we elucidated effect of Cu2+ on the energy producing metabolic enzyme creatine kinase (CK), which might directly affect energy metabolism and homeostasis of P. sinensis. Method: We first conducted molecular dynamics (MD) simulations between P-CK and Cu2+ and conducted the inactivation kinetics including spectrofluorimetry study. Results: MD simulation showed that Cu2+ blocked the binding site of the ATP cofactor, indicating that Cu2+ could directly inactivate P-CK. We prepared the muscle type of CK (P-CK) and confirmed that Cu2+ conspicuously inactivated the activity of P-CK (IC50 = 24.3 ?M) and exhibited non-competitive inhibition manner with creatine and ATP in a first-order kinetic process. This result was well matched to the MD simulation results that Cu2+-induced non-competitive inactivation of P-CK. The spectrofluorimetry study revealed that Cu2+ induced tertiary structure changes in PCK accompanying with the exposure of hydrophobic surfaces. Interestingly, the addition of osmolytes (glycine, proline, and liquaemin) effectively restored activity of the Cu2+-inactivated P-CK. Conclusion: Our study illustrates the Cu2+-mediated unfolding of P-CK with disruption of the enzymatic function and the protective restoration role of osmolytes on P-CK inactivation. This study provides information of interest on P-CK as a metabolic enzyme of ectothermic animal in response to Cu2+ binding.

ISSN 

0929-8665

Link 

http://dx.doi.org/10.2174/0929866524666170227122706

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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