SPR Analysis of Adrenodoxin Interaction with CYP11B1 Embedded into the Lipid Bilayer

IF 1.1 Q4 CELL BIOLOGY

Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology Pub Date : 2025-04-24 DOI:10.1134/S1990747824700466

O. V. Gnedenko, E. O. Yablokov, L. A. Kaluzhskiy, Y. V. Mezentsev, T. V. Tsybruk, M. A. Karpova, A. S. Ivanov

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Abstract

Surface plasmon resonance (SPR) is a relevant method for studying interactions involving membrane proteins, mitochondrial proteins in particular. Currently, there is only a little information on the interaction of mitochondrial proteins with their partners, such as electron carriers, in lipid microenvironment. In this work, using cytochrome P450 11β-hydroxylase (CYP11B1) and adrenodoxin (Adx) as an example, we showed that SPR analysis can be used for studying protein–protein interactions in lipid microenvironment. Kinetic and equilibrium parameters of the CYP11B1-Adx complexes in lipid bilayers of different compositions were determined. Our results reveal that the CYP11B1-Adx complex in cardiolipin-containing bilayer demonstrates increased stability: nearly tenfold reduction in the equilibrium dissociation constant (Kd) compared to the bilayer without cardiolipin. The interaction between redox proteins is highly sensitive to the lipid structure. The immobilized protein and the lipid composition should be selected in each case, but overall, the obtained data are highly complementary for studying membrane proteins.

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肾上腺素还毒素与嵌入脂质双分子层的CYP11B1相互作用的SPR分析

表面等离子体共振（SPR）是研究膜蛋白，特别是线粒体蛋白相互作用的一种相关方法。目前，关于线粒体蛋白与其伴侣（如电子载体）在脂质微环境中的相互作用的信息很少。在本研究中，我们以细胞色素P450 11β-羟化酶（CYP11B1）和肾上腺素还毒素（Adx）为例，证明了SPR分析可以用于研究脂质微环境中蛋白-蛋白相互作用。测定了不同组成的脂质双分子层中CYP11B1-Adx复合物的动力学和平衡参数。我们的研究结果表明，在含有心磷脂的双分子层中，CYP11B1-Adx复合物表现出更高的稳定性：与不含心磷脂的双分子层相比，平衡解离常数（Kd）降低了近10倍。氧化还原蛋白之间的相互作用对脂质结构高度敏感。每种情况下都需要选择固定蛋白和脂质组成，但总的来说，所获得的数据对于研究膜蛋白具有很强的互补性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology CELL BIOLOGY-

CiteScore

1.40

自引率

0.00%

发文量

期刊介绍： Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology is an international peer reviewed journal that publishes original articles on physical, chemical, and molecular mechanisms that underlie basic properties of biological membranes and mediate membrane-related cellular functions. The primary topics of the journal are membrane structure, mechanisms of membrane transport, bioenergetics and photobiology, intracellular signaling as well as membrane aspects of cell biology, immunology, and medicine. The journal is multidisciplinary and gives preference to those articles that employ a variety of experimental approaches, basically in biophysics but also in biochemistry, cytology, and molecular biology. The journal publishes articles that strive for unveiling membrane and cellular functions through innovative theoretical models and computer simulations.