AFM study of the interaction of cytochrome P450 2C9 with phospholipid bilayers

Nussio, Matthew R.; Voelcker, Nicolas H.; Miners, John O.; Lewis, Benjamin C.; Sykes, Matthew J.; Shapter, Joseph G.

doi:10.1016/j.chemphyslip.2009.11.003

Cited by 18 publications

(15 citation statements)

References 68 publications

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“…Carbofuran induced the phospholipid in the membrane of hepatocytes which might be involved in activating a signaling cascade, including protein kinase C (PKC) or heat shock protein 70 (HSP70) [38]. The role of phospholipid in the activation of enzymatic activity of membrane-bound protein, including cytochrome P450 has been studied by many investigators [7], [8], [10], [39], [40], [41]. Many phospholipids, such as, dilaurylphosphatidylcholine (DLPC) and dilaurylphosphatidylserine (DLPS) have been used to reconstitute the enzymatic activity of cytochrome P450 protein [3], [33], [41], [42].…”

Section: Discussionmentioning

confidence: 99%

Membrane Phospholipid Augments Cytochrome P4501a Enzymatic Activity by Modulating Structural Conformation during Detoxification of Xenobiotics

Ghosh

Ray

2013

PLoS ONE

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Cytochrome P450 is a superfamily of membrane-bound hemoprotein that gets involved with the degradation of xenobiotics and internal metabolites. Accumulated body of evidence indicates that phospholipids play a crucial role in determining the enzymatic activity of cytochrome P450 in the microenvironment by modulating its structure during detoxification; however, the structure-function relationship of cytochrome P4501A, a family of enzymes responsible for degrading lipophilic aromatic hydrocarbons, is still not well defined. Inducibility of cytochrome P4501A in cultured catfish hepatocytes in response to carbofuran, a widely used pesticide around the world, was studied earlier in our laboratory. In this present investigation, we observed that treating catfish with carbofuran augmented total phospholipid in the liver. We examined the role of phospholipid on the of cytochrome P4501A-marker enzyme which is known as ethoxyresorufin-O-deethylase (EROD) in the context of structure and function. We purified the carbofuran-induced cytochrome P4501A protein from catfish liver. Subsequently, we examined the enzymatic activity of purified P4501A protein in the presence of phospholipid, and studied how the structure of purified protein was influenced in the phospholipid environment. Membrane phospholipid appeared to accelerate the enzymatic activity of EROD by changing its structural conformation and thus controlling the detoxification of xenobiotics. Our study revealed the missing link of how the cytochrome P450 restores its enzymatic activity by changing its structural conformation in the phospholipid microenvironment.

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Section: Discussionmentioning

confidence: 99%

Membrane Phospholipid Augments Cytochrome P4501a Enzymatic Activity by Modulating Structural Conformation during Detoxification of Xenobiotics

Ghosh

Ray

2013

PLoS ONE

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“…This has been shown by experiments that probed the recognition of reconstituted microsomes by site-directed antibodies against peptides of the human CYP2B1 and the rabbit CYP2B4 [2], [8]. Engineered CYPs with the transmembrane domain removed retain membrane-binding properties [2], [9]. The height of the globular domain above the lipid bilayer has been estimated to be 35±9 Å [10].…”

Section: Introductionmentioning

confidence: 97%

Structure and Dynamics of the Membrane-Bound Cytochrome P450 2C9

et al. 2011

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The microsomal, membrane-bound, human cytochrome P450 (CYP) 2C9 is a liver-specific monooxygenase essential for drug metabolism. CYPs require electron transfer from the membrane-bound CYP reductase (CPR) for catalysis. The structural details and functional relevance of the CYP-membrane interaction are not understood. From multiple coarse grained molecular simulations started with arbitrary configurations of protein-membrane complexes, we found two predominant orientations of CYP2C9 in the membrane, both consistent with experiments and conserved in atomic-resolution simulations. The dynamics of membrane-bound and soluble CYP2C9 revealed correlations between opening and closing of different tunnels from the enzyme's buried active site. The membrane facilitated the opening of a tunnel leading into it by stabilizing the open state of an internal aromatic gate. Other tunnels opened selectively in the simulations of product-bound CYP2C9. We propose that the membrane promotes binding of liposoluble substrates by stabilizing protein conformations with an open access tunnel and provide evidence for selective substrate access and product release routes in mammalian CYPs. The models derived here are suitable for extension to incorporate other CYPs for oligomerization studies or the CYP reductase for studies of the electron transfer mechanism, whereas the modeling procedure is generally applicable to study proteins anchored in the bilayer by a single transmembrane helix.

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“…Метод атомно-силовой микроскопии применяется в качестве контроля мономерного состояния цитохромов P450 при иммобилизации на поверхности биосенсоров [126], а также для определения оптимального модификатора поверхности, в том числе при совместной иммобилизации с белками-партнёрами [127,128], CYP17A1 и CYP19A1 [129].…”

Section: метод атомно-силовой микроскопииunclassified

Methods for determining of cytochrome P450 isozymes functional activity

Kuzikov

Masamrekh

Archakov³

et al. 2018

BIOMED KHIM

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The review is dedicated to modern methods and technologies for determining of cytochrome P450 isozymes functional activity, such as absorbance and fluorescent spectroscopy, electron paramagnetic resonance (EPR), nuclear magnetic resonance (NMR), Raman, Mossbauer, and X-ray spectroscopy, surface plasmon resonance (SPR), atomic force microscopy (AFM). Methods of molecular genetic analysis were reviewed from personalized medicine point of view. The use of chromate-mass-spectrometric methods for cytochrome P450-dependent catalytic reactions' products was discussed. The review covers modern electrochemical systems based on cytochrome P450 isozymes for their catalytic activity analysis, their use in practice and further development perspectives for experimental pharmacology, biotechnology and translational medicine.

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AFM study of the interaction of cytochrome P450 2C9 with phospholipid bilayers

Cited by 18 publications

References 68 publications

Membrane Phospholipid Augments Cytochrome P4501a Enzymatic Activity by Modulating Structural Conformation during Detoxification of Xenobiotics

Membrane Phospholipid Augments Cytochrome P4501a Enzymatic Activity by Modulating Structural Conformation during Detoxification of Xenobiotics

Structure and Dynamics of the Membrane-Bound Cytochrome P450 2C9

Methods for determining of cytochrome P450 isozymes functional activity

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