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Impact of Lipid Matrix Composition on Activity of Membranotropic Enzymes Galactonolactone Oxidase from Trypanosoma cruzi and L-Galactono-1,4-Lactone Dehydrogenase from Arabidopsis thaliana in the System of Reverse Micelles

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Abstract

The study of many membrane enzymes in an aqueous medium is difficult due to the loss of their catalytic activity, which makes it necessary to use membrane-like systems, such as reverse micelles of surfactants in nonpolar organic solvents. However, it should be taken into account that the micelles are a simplified model of natural membranes, since membranes contain many different components, a significant part of which are phospholipids. In this work, we studied impact of the main phospholipids, phosphatidylcholine (PC) and phosphatidylethanolamine (PE), on activity of the membrane enzymes using galactonolactone oxidase from Trypanosoma cruzi (TcGAL) and L-galactono-1,4-lactone dehydrogenase from Arabidopsis thaliana (AtGALDH) as examples. Effect of the structure (and charge) of the micelle-forming surfactant itself on the activity of both enzymes has been studied using an anionic surfactant (AOT), a neutral surfactant (Brij-96), and a mixture of cationic and anionic surfactants (CTAB and AOT) as examples. The pronounced effect of addition of PC and PE lipids on the activity of AtGALDH and TcGAL has been detected, which manifests as increase in catalytic activity and significant change in the activity profile. This can be explained by formation of the tetrameric form of enzymes and/or protein–lipid complexes. By varying composition and structure of the micelle-forming surfactants (AOT, CTAB, and Brij-96) it has been possible to change catalytic properties of the enzyme due to effect of the surfactant on the micelle size, lipid mobility, charge, and rigidity of the matrix itself.

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Abbreviations

AL:

D-arabinono-1,4-lactone

AOT:

bis-2-ethylhexyl sulfosuccinate, anionic SAA

AtGALDH:

L-galactono-1,4-lactone dehydrogenase from Arabidopsis thaliana

CTAB:

cetyltrimethylammonium bromide, cationic SAA

DCPIP:

2,6-dichlorophenolindophenol

EA:

electron acceptor

GL:

L-galactono-1,4-lactone

PC:

phosphatidylcholine

PE:

phosphatidylethanolamine

PMS:

phenazine methosulfate

SAA:

surface active agents

TcGAL:

galactonolactone oxidase from Trypanosoma cruzi

W0 :

the hydration degree

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Acknowledgments

Equipment acquired through the program of Moscow State University Development, FT-IR Bruker Tensor 27 spectrometer (Germany) and CD-spectrometer Jasco J-815 (JASCO, Japan), was used in this work.

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  1. Faculty of Chemistry, Lomonosov Moscow State University, 119991, Moscow, Russia

    Andrey A. Chudin & Elena V. Kudryashova

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  1. Andrey A. Chudin
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Contributions

E.V.K. concept of the study and supervision of the work; A.A.Ch. conducting experiments; E.V.K. and A.A.Ch. discussion of the study results; A.A.Ch. writing text of the paper; E.V.K. and A.A.Ch. editing text of the paper.

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Correspondence to Elena V. Kudryashova.

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Chudin, A.A., Kudryashova, E.V. Impact of Lipid Matrix Composition on Activity of Membranotropic Enzymes Galactonolactone Oxidase from Trypanosoma cruzi and L-Galactono-1,4-Lactone Dehydrogenase from Arabidopsis thaliana in the System of Reverse Micelles. Biochemistry Moscow 88, 2073–2083 (2023). https://doi.org/10.1134/S0006297923120106

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