Protein oligomerization induced by oleic acid at the solid-liquid interface–equine lysozyme cytotoxic complexes.
Abstract: Protein oligomeric complexes have emerged as a major target of current research because of their key role in aggregation processes in living systems and in vitro. Hydrophobic and charged surfaces may favour the self-assembly process by recruiting proteins and modifying their interactions. We found that equine lysozyme assembles into multimeric complexes with oleic acid (ELOA) at the solid-liquid interface within an ion-exchange chromatography column preconditioned with oleic acid. The properties of ELOA were characterized using NMR, spectroscopic methods and atomic force microscopy, and showed similarity with both amyloid oligomers and the complexes with oleic acid and its structural homologous protein alpha-lactalbumin, known as human alpha-lactalbumin made lethal for tumour cells (HAMLET). As determined by NMR diffusion measurements, ELOA may consist of 4-30 lysozyme molecules. Each lysozyme molecule is able to bind 11-48 oleic acids in various preparations. Equine lysozyme acquired a partially unfolded conformation in ELOA, as evident from its ability to bind hydrophobic dye 8-anilinonaphthalene-1-sulfonate. CD and NMR spectra. Similar to amyloid oligomers, ELOA also interacts with thioflavin-T dye, shows a spherical morphology, assembles into ring-shaped structures, as monitored by atomic force microscopy, and exerts a toxic effect in cells. Studies of well-populated ELOA shed light on the nature of the amyloid oligomers and HAMLET complexes, suggesting that they constitute one large family of cytotoxic proteinaceous species. The hydrophobic surfaces can be used profitably to produce complexes with very distinct properties compared to their precursor proteins.
Publication Date: 2009-07-07 PubMed ID: 19594832DOI: 10.1111/j.1742-4658.2009.07107.xGoogle Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research studied the formation of protein complexes, specifically equine lysozyme with oleic acid (ELOA), and their relevance in biological aggregation processes and implied cytotoxicity. These complexes show similarities with amyloid oligomers and HAMLET protein complexes, establishing a possible link between these different protein structures.
Study Focus and Methodology
- The researchers focused on protein oligomeric complexes, mainly as they play a crucial role within aggregation processes in biological systems and in laboratory conditions.
- The experiment centered on the assembly of equine lysozyme, a protein, into larger complexes with oleic acid. The assembly took place at the solid-liquid interface within an ion-exchange chromatography column that had been preconditioned with oleic acid.
Findings and Characterization
- The created complexes, termed ELOA (Equine Lysozyme-Oleic Acid), were analyzed using NMR spectroscopy, other spectroscopic methods, and atomic force microscopy.
- The structure of ELOA showed similarities to amyloid oligomers and to complexes known as HAMLET (human alpha-lactalbumin made lethal for tumour cells), which form between oleic acid and a structurally similar protein to lysozyme.
- The ELOA was estimated to contain between 4 and 30 lysozyme molecules, each binding to between 11 and 48 oleic acid molecules.
- Within the ELOA complex, it was observed that the equine lysozyme adopted a partially unfolded formation which allowed it to bind to hydrophobic dye 8-anilinonaphthalene-1-sulfonate, as shown through CD and NMR spectra.
- The ELOA complex mirrored amyloid oligomers in its interaction with thioflavin-T dye and exhibited similar spherical morphology and ring-shaped structures.
Implications and Conclusions
- ELOA exerted a toxic effect on cells, thereby demonstrating potential cytotoxic attributes, much like the amyloid oligomers and HAMLET complexes.
- The properties and cytotoxic implications of the well-populated ELOA complex offered insights into the nature of amyloid oligomers and HAMLET complexes, implying that these diverse entities may belong to a single large family of cytotoxic proteins.
- The researchers highlighted the potential to use hydrophobic surfaces to generate complexes with distinct properties compared to their precursor proteins, which could be a valuable tool in future research or applications.
Cite This Article
APA
Wilhelm K, Darinskas A, Noppe W, Duchardt E, Mok KH, Vukojević V, Schleucher J, Morozova-Roche LA.
(2009).
Protein oligomerization induced by oleic acid at the solid-liquid interface–equine lysozyme cytotoxic complexes.
FEBS J, 276(15), 3975-3989.
https://doi.org/10.1111/j.1742-4658.2009.07107.x Publication
Researcher Affiliations
- Department of Medical Biochemistry and Biophysics, Umeå University, Sweden.
MeSH Terms
- Animals
- Cytotoxins / chemistry
- Cytotoxins / metabolism
- Fluorescent Dyes
- Horses
- Humans
- Kinetics
- Lactalbumin / chemistry
- Lactalbumin / metabolism
- Magnetic Resonance Spectroscopy
- Microscopy, Atomic Force
- Muramidase / chemistry
- Muramidase / metabolism
- Oleic Acids / chemistry
- Oleic Acids / metabolism
- Spectrophotometry
Citations
This article has been cited 13 times.- El-Fakharany EM, Redwan EM. Protein-lipid complexes: molecular structure, current scenarios and mechanisms of cytotoxicity.. RSC Adv 2019 Nov 11;9(63):36890-36906.
- Chetta KE, Alcorn JL, Baatz JE, Wagner CL. Cytotoxic Lactalbumin-Oleic Acid Complexes in the Human Milk Diet of Preterm Infants.. Nutrients 2021 Nov 30;13(12).
- Elizarova A, Sokolov A, Kostevich V, Kisseleva E, Zelenskiy E, Zakharova E, Panasenko O, Budevich A, Semak I, Egorov V, Pontarollo G, De Filippis V, Vasilyev V. Interaction of Lactoferrin with Unsaturated Fatty Acids: In Vitro and In Vivo Study of Human Lactoferrin/Oleic Acid Complex Cytotoxicity.. Materials (Basel) 2021 Mar 25;14(7).
- Frislev HS, Nielsen J, Nylandsted J, Otzen D. Using Liprotides to Deliver Cholesterol to the Plasma Membrane.. J Membr Biol 2018 Aug;251(4):581-592.
- Kataev A, Zherelova O, Grishchenko V. A Characeae Cells Plasma Membrane as a Model for Selection of Bioactive Compounds and Drugs: Interaction of HAMLET-Like Complexes with Ion Channels of Chara corallina Cells Plasmalemma.. J Membr Biol 2016 Dec;249(6):801-811.
- Woods KN, Pfeffer J. Using THz Spectroscopy, Evolutionary Network Analysis Methods, and MD Simulation to Map the Evolution of Allosteric Communication Pathways in c-Type Lysozymes.. Mol Biol Evol 2016 Jan;33(1):40-61.
- Clementi EA, Wilhelm KR, Schleucher J, Morozova-Roche LA, Hakansson AP. A complex of equine lysozyme and oleic acid with bactericidal activity against Streptococcus pneumoniae.. PLoS One 2013;8(11):e80649.
- Ho JC, Storm P, Rydström A, Bowen B, Alsin F, Sullivan L, Ambite I, Mok KH, Northen T, Svanborg C. Lipids as tumoricidal components of human α-lactalbumin made lethal to tumor cells (HAMLET): unique and shared effects on signaling and death.. J Biol Chem 2013 Jun 14;288(24):17460-71.
- Nakamura T, Aizawa T, Kariya R, Okada S, Demura M, Kawano K, Makabe K, Kuwajima K. Molecular mechanisms of the cytotoxicity of human α-lactalbumin made lethal to tumor cells (HAMLET) and other protein-oleic acid complexes.. J Biol Chem 2013 May 17;288(20):14408-14416.
- Storm P, Klausen TK, Trulsson M, Ho C S J, Dosnon M, Westergren T, Chao Y, Rydström A, Yang H, Pedersen SF, Svanborg C. A unifying mechanism for cancer cell death through ion channel activation by HAMLET.. PLoS One 2013;8(3):e58578.
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- Bruhn O, Grötzinger J, Cascorbi I, Jung S. Antimicrobial peptides and proteins of the horse--insights into a well-armed organism.. Vet Res 2011 Sep 2;42(1):98.
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