Assessing the Interactions of Statins with Human Adenylate Kinase Isoenzyme 1: Fluorescence and Enzyme Kinetic Studies.

Magdalena Wujak,Anna Kozakiewicz,Zuzanna Sokolowska,Marcin Budny,Andrzej Wojtczak,Anna Ciarkowska,Joanna I Loch,Magdalena Barwiolek

doi:10.3390/ijms22115541

Magdalena Wujak, Anna Kozakiewicz + Show 6 more

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https://doi.org/10.3390/ijms22115541

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Abstract

Statins are the most effective cholesterol-lowering drugs. They also exert many pleiotropic effects, including anti-cancer and cardio- and neuro-protective. Numerous nano-sized drug delivery systems were developed to enhance the therapeutic potential of statins. Studies on possible interactions between statins and human proteins could provide a deeper insight into the pleiotropic and adverse effects of these drugs. Adenylate kinase (AK) was found to regulate HDL endocytosis, cellular metabolism, cardiovascular function and neurodegeneration. In this work, we investigated interactions between human adenylate kinase isoenzyme 1 (hAK1) and atorvastatin (AVS), fluvastatin (FVS), pravastatin (PVS), rosuvastatin (RVS) and simvastatin (SVS) with fluorescence spectroscopy. The tested statins quenched the intrinsic fluorescence of hAK1 by creating stable hAK1-statin complexes with the binding constants of the order of 104 M−1. The enzyme kinetic studies revealed that statins inhibited hAK1 with significantly different efficiencies, in a noncompetitive manner. Simvastatin inhibited hAK1 with the highest yield comparable to that reported for diadenosine pentaphosphate, the only known hAK1 inhibitor. The determined AK sensitivity to statins differed markedly between short and long type AKs, suggesting an essential role of the LID domain in the AK inhibition. Our studies might open new horizons for the development of new modulators of short type AKs.

Highlights

Introduction iationsStatins are potent competitive inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase
Differences in the structure of the LID domain are the basis for the Adenylate kinase (AK) classification into short and long type AKs, which was related to different inhibitory efficiencies of diadenosine polyphosphates as inhibitors
Recombinant human adenylate kinase and adenylate kinase from Geobacillus stearothermophilus (AKst, long type AK) were produced in the bacterial expression system based on the E. coli strain BL21CodonPlus (DE3)-RIL and pET-3a(+) expression vector

Summary

Introduction

Statins are potent competitive inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase. They inhibit the conversion of HMG-CoA to mevalonate, which is the rate-limiting step in the cholesterol biosynthesis [1]. For their ability to enhance the clearance of plasma low-density lipoproteins, statins have become the first-line therapy in the prevention of atherosclerotic cardiovascular disease [1,2]. These drugs possess a wide spectrum of so-called pleiotropic (cholesterol-independent) effects that may largely contribute to clinical benefits of the statin therapy and expand their use to treat other diseases [2]. The protective effects of statins on the vascular endothelium are related to the enhanced production and bioavailability of nitric oxide (NO) as well as restoration

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