scholarly journals Nonequilibrium fluctuations of lipid membranes by the rotating motor protein F1F0-ATP synthase

2017 ◽  
Vol 114 (43) ◽  
pp. 11291-11296 ◽  
Author(s):  
Víctor G. Almendro-Vedia ◽  
Paolo Natale ◽  
Michael Mell ◽  
Stephanie Bonneau ◽  
Francisco Monroy ◽  
...  

ATP synthase is a rotating membrane protein that synthesizes ATP through proton-pumping activity across the membrane. To unveil the mechanical impact of this molecular active pump on the bending properties of its lipid environment, we have functionally reconstituted the ATP synthase in giant unilamellar vesicles and tracked the membrane fluctuations by means of flickering spectroscopy. We find that ATP synthase rotates at a frequency of about 20 Hz, promoting large nonequilibrium deformations at discrete hot spots in lipid vesicles and thus inducing an overall membrane softening. The enhanced nonequilibrium fluctuations are compatible with an accumulation of active proteins at highly curved membrane sites through a curvature−protein coupling mechanism that supports the emergence of collective effects of rotating ATP synthases in lipid membranes.

Processes ◽  
2021 ◽  
Vol 9 (6) ◽  
pp. 945
Author(s):  
Qiong Wang ◽  
Ning Hu ◽  
Jincan Lei ◽  
Qiurong Qing ◽  
Jing Huang ◽  
...  

Lipid vesicles, especially giant lipid vesicles (GLVs), are usually adopted as cell membrane models and their preparation has been widely studied. However, the effects of some nonelectrolytes on GLV formation have not been specifically studied so far. In this paper, the effects of the nonelectrolytes, including sucrose, glucose, sorbitol and ethanol, and their coexistence with sodium chloride, on the lipid hydration and GLV formation were investigated. With the hydration method, it was found that the sucrose, glucose and sorbitol showed almost the same effect. Their presence in the medium enhanced the hydrodynamic force on the lipid membranes, promoting the GLV formation. GLV formation was also promoted by the presence of ethanol with ethanol volume fraction in the range of 0 to 20 percent, but higher ethanol content resulted in failure of GLV formation. However, the participation of sodium chloride in sugar solution and ethanol solution stabilized the lipid membranes, suppressing the GLV formation. In addition, the ethanol and the sodium chloride showed the completely opposite effects on lipid hydration. These results could provide some suggestions for the efficient preparation of GLVs.


1988 ◽  
Vol 104 (2) ◽  
pp. 179-191 ◽  
Author(s):  
B. Christensen ◽  
M. Gutweiler ◽  
E. Grell ◽  
N. Wagner ◽  
R. Pabst ◽  
...  

1996 ◽  
Vol 318 (1) ◽  
pp. 343-349 ◽  
Author(s):  
Susan K BUCHANAN ◽  
John E. WALKER

A new chromatographic procedure has been developed for the isolation of F1Fo-ATPase and NADH:ubiquinone oxidoreductase (complex I) from a single batch of bovine heart mitochondria. The method employed dodecyl β-Δ-maltoside, a monodisperse, homogeneous detergent in which many respiratory complexes exhibit high activity, for solubilization and subsequent purification by ammonium sulphate fractionation and column chromatography. A combination of anion-exchange, gel-filtration, and dye-ligand affinity chromatography was used to purify both complexes to homogeneity. The F1Fo-ATPase preparation contains only the 16 known subunits of the enzyme. It has oligomycin-sensitive ATP hydrolysis activity and, as demonstrated elsewhere, when reconstituted into lipid vesicles it is capable of ATP-dependent proton pumping and of ATP synthesis driven by a proton gradient [Groth and Walker (1996) Biochem. J. 318, 351–357]. The complex I preparation contains all of the subunits identified in other preparations of the enzyme, and has rotenone-sensitive NADH:ubiquinone oxidoreductase and NADH:ferricyanide oxidoreductase activities. The procedure is rapid and reproducible, yielding 50–80 mg of purified F1Fo-ATPase and 20–40 mg of purified complex I from 1 g of mitochondrial membranes. Both preparations are devoid of phospholipids, and gel filtration and dynamic light scattering experiments indicate that they are monodisperse. Therefore, the preparations fulfil important prerequisites for structural analysis.


Genetics ◽  
2001 ◽  
Vol 159 (3) ◽  
pp. 929-938
Author(s):  
G D Clark-Walker ◽  
X J Chen

Abstract Loss of mtDNA or mitochondrial protein synthesis cannot be tolerated by wild-type Kluyveromyces lactis. The mitochondrial function responsible for ρ0-lethality has been identified by disruption of nuclear genes encoding electron transport and F0-ATP synthase components of oxidative phosphorylation. Sporulation of diploid strains heterozygous for disruptions in genes for the two components of oxidative phosphorylation results in the formation of nonviable spores inferred to contain both disruptions. Lethality of spores is thought to result from absence of a transmembrane potential, ΔΨ, across the mitochondrial inner membrane due to lack of proton pumping by the electron transport chain or reversal of F1F0-ATP synthase. Synergistic lethality, caused by disruption of nuclear genes, or ρ0-lethality can be suppressed by the atp2.1 mutation in the β-subunit of F1-ATPase. Suppression is viewed as occurring by an increased hydrolysis of ATP by mutant F1, allowing sufficient electrogenic exchange by the translocase of ADP in the matrix for ATP in the cytosol to maintain ΔΨ. In addition, lethality of haploid strains with a disruption of AAC encoding the ADP/ATP translocase can be suppressed by atp2.1. In this case suppression is considered to occur by mutant F1 acting in the forward direction to partially uncouple ATP production, thereby stimulating respiration and relieving detrimental hyperpolarization of the inner membrane. Participation of the ADP/ATP translocase in suppression of ρ0-lethality is supported by the observation that disruption of AAC abolishes suppressor activity of atp2.1.


2016 ◽  
Vol 160 (4) ◽  
pp. 205-215 ◽  
Author(s):  
Madhavan Narayanan ◽  
Joseph A. Sakyiama ◽  
Mahmoud M. Elguindy ◽  
Eiko Nakamaru-Ogiso

2013 ◽  
Vol 33 (5) ◽  
Author(s):  
Chi L. L. Pham ◽  
Roberto Cappai

The deposition of α-syn (α-synuclein) as amyloid fibrils and the selective loss of DA (dopamine) containing neurons in the substantia nigra are two key features of PD (Parkinson's disease). α-syn is a natively unfolded protein and adopts an α-helical conformation upon binding to lipid membrane. Oligomeric species of α-syn have been proposed to be the pathogenic species associated with PD because they can bind lipid membranes and disrupt membrane integrity. DA is readily oxidized to generate reactive intermediates and ROS (reactive oxygen species) and in the presence of DA, α-syn form of SDS-resistant soluble oligomers. It is postulated that the formation of the α-syn:DA oligomers involves the cross-linking of DA-melanin with α-syn, via covalent linkage, hydrogen and hydrophobic interactions. We investigate the effect of lipids on DA-induced α-syn oligomerization and studied the ability of α-syn:DA oligomers to interact with lipids vesicles. Our results show that the interaction of α-syn with lipids inhibits the formation of DA-induced α-syn oligomers. Moreover, the α-syn:DA oligomer cannot interact with lipid vesicles or cause membrane permeability. Thus, the formation of α-syn:DA oligomers may alter the actions of α-syn which require membrane association, leading to disruption of its normal cellular function.


1997 ◽  
Vol 489 ◽  
Author(s):  
Christian W. Maier ◽  
Almuth Behrisch ◽  
Annette Kloboucek ◽  
Rudolf Merkel

AbstractWe used the micropipet aspiration technique for a study of biomembrane adhesion. Adhesion was caused by contact site A, a highly specific cell adhesion molecule, reconstituted in lipid vesicles of DOPC with 5 %(mol/mol) DOPE-PEG2000. We found adhesion and subsequent receptor aggregation in the contact zone. Additionally, electrostatic modulation of membrane adhesion was studied. Whereas addition of the negatively charged lipid SOPS to the lecithin (SOPC) host membrane suppressed adhesion due to electrostatic repulsion, a positively charged lipid (DOTAP) was surprisingly ineffective. This might be due to either phase separation of the mixture or DOTAP changing other membrane properties as bending stiffness and the Hamaker constant.


2017 ◽  
Vol 114 (6) ◽  
pp. E1009-E1017 ◽  
Author(s):  
Michele Perni ◽  
Céline Galvagnion ◽  
Alexander Maltsev ◽  
Georg Meisl ◽  
Martin B. D. Müller ◽  
...  

The self-assembly of α-synuclein is closely associated with Parkinson’s disease and related syndromes. We show that squalamine, a natural product with known anticancer and antiviral activity, dramatically affects α-synuclein aggregation in vitro and in vivo. We elucidate the mechanism of action of squalamine by investigating its interaction with lipid vesicles, which are known to stimulate nucleation, and find that this compound displaces α-synuclein from the surfaces of such vesicles, thereby blocking the first steps in its aggregation process. We also show that squalamine almost completely suppresses the toxicity of α-synuclein oligomers in human neuroblastoma cells by inhibiting their interactions with lipid membranes. We further examine the effects of squalamine in a Caenorhabditis elegans strain overexpressing α-synuclein, observing a dramatic reduction of α-synuclein aggregation and an almost complete elimination of muscle paralysis. These findings suggest that squalamine could be a means of therapeutic intervention in Parkinson’s disease and related conditions.


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