Protein oligomerization induced by oleic acid at the solid-liquid interface--equine lysozyme cytotoxic complexes.
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.
Wilhelm
Kristina
K
Department of Medical Biochemistry and Biophysics, Umeå University, Sweden.
Darinskas
Adas
A
Noppe
Wim
W
Duchardt
Elke
E
Mok
K Hun
KH
Vukojević
Vladana
V
Schleucher
Jürgen
J
Morozova-Roche
Ludmilla A
LA
D000818:Animals; D003603:Cytotoxins / Q000737:chemistry / Q000378:metabolism; D005456:Fluorescent Dyes; D006736:Horses; D006801:Humans; D007700:Kinetics; D007768:Lactalbumin / Q000737:chemistry / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D018625:Microscopy, Atomic Force; D009113:Muramidase / Q000737:chemistry / Q000378:metabolism*; D009829:Oleic Acids / Q000737:chemistry / Q000378:metabolism*; D013053:Spectrophotometry
D003603:Cytotoxins; D005456:Fluorescent Dyes; C506422:HAMLET complex, human; D009829:Oleic Acids; D007768:Lactalbumin; D009113:Muramidase