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1.
J Membr Biol ; 252(2-3): 115-130, 2019 06.
Article in English | MEDLINE | ID: mdl-30877332

ABSTRACT

Of all the macromolecular assemblies of life, the least understood is the biomembrane. This is especially true in regard to its atomic structure. Ideas on biomembranes, developed in the last 200 years, culminated in the fluid mosaic model of the membrane. In this essay, I provide a historical outline of how we arrived at our current understanding of biomembranes and the models we use to describe them. A selection of direct experimental findings on the nano-scale structure of biomembranes is taken up to discuss their physical nature, and special emphasis is put on the surprising insights that arise from atomic scale descriptions.


Subject(s)
Cell Membrane/ultrastructure , Membrane Lipids/chemistry , Membrane Microdomains/ultrastructure , Membrane Proteins/ultrastructure , Cell Membrane/metabolism , Crystallography, X-Ray , Enterococcus hirae/metabolism , Enterococcus hirae/ultrastructure , Eukaryotic Cells/metabolism , Eukaryotic Cells/ultrastructure , Halobacterium salinarum/metabolism , Halobacterium salinarum/ultrastructure , Humans , Hydrophobic and Hydrophilic Interactions , Membrane Lipids/metabolism , Membrane Microdomains/metabolism , Membrane Proteins/metabolism , Models, Biological , Protein Conformation
2.
Sci Rep ; 8(1): 15632, 2018 10 23.
Article in English | MEDLINE | ID: mdl-30353110

ABSTRACT

EhV-ATPase is an ATP-driven Na+ pump in the eubacteria Enterococcus hirae (Eh). Here, we present the first entire structure of detergent-solubilized EhV-ATPase by single-particle cryo-electron microscopy (cryo-EM) using Zernike phase plate. The cryo-EM map dominantly showed one of three catalytic conformations in this rotary enzyme. To further stabilize the originally heterogeneous structure caused by the ATP hydrolysis states of the V1-ATPases, a peptide epitope tag system was adopted, in which the inserted peptide epitope sequence interfered with rotation of the central rotor by binding the Fab. As a result, the map unexpectedly showed another catalytic conformation of EhV-ATPase. Interestingly, these two conformations identified with and without Fab conversely coincided with those of the minor state 2 and the major state 1 of Thermus thermophilus V/A-ATPase, respectively. The most prominent feature in EhV-ATPase was the off-axis rotor, where the cytoplasmic V1 domain was connected to the transmembrane Vo domain through the off-axis central rotor. Furthermore, compared to the structure of ATP synthases, the larger size of the interface between the transmembrane a-subunit and c-ring of EhV-ATPase would be more advantageous for active ion pumping.


Subject(s)
Cryoelectron Microscopy , Enterococcus hirae/enzymology , Vacuolar Proton-Translocating ATPases/ultrastructure , Bacterial Proteins/metabolism , Bacterial Proteins/ultrastructure , Cryoelectron Microscopy/methods , Enterococcus hirae/metabolism , Enterococcus hirae/ultrastructure , Gram-Positive Bacterial Infections/microbiology , Humans , Models, Molecular , Protein Subunits/metabolism , Recombinant Proteins/metabolism , Recombinant Proteins/ultrastructure , Vacuolar Proton-Translocating ATPases/metabolism
3.
Lett Appl Microbiol ; 65(3): 220-225, 2017 Sep.
Article in English | MEDLINE | ID: mdl-28609553

ABSTRACT

A low-intensity electromagnetic field of extremely high frequency has inhibitory and stimulatory effects on bacteria, including Enterococcus hirae. It was shown that the low-intensity (the incident power density of 0·06 mW cm-2 ) electromagnetic field at the frequencies of 51·8 GHz and 53 GHz inhibited E. hirae ATCC 9790 bacterial growth rate; a stronger effect was observed with 53 GHz, regardless of exposure duration (0·5 h, 1 h or 2 h). Scanning electron microscopy analysis of these effects has been done; the cells were of spherical shape. Electromagnetic field at 53 GHz, but not 51·8 GHz, changed the cell size-the diameter was enlarged 1·3 fold at 53 GHz. These results suggest the difference in mechanisms of action on bacteria for electromagnetic fields at 51·8 GHz and 53 GHz. SIGNIFICANCE AND IMPACT OF THE STUDY: A stronger inhibitory effect of low-intensity electromagnetic field on Enterococcus hirae ATCC 9790 bacterial growth rate was observed with 53 GHz vs 51·8 GHz, regardless of exposure duration. Scanning electron microscopy analysis showed that almost all irradiated cells in the population have spherical shapes similar to nonirradiated ones, but they have increased diameters in case of irradiated cells at 53 GHz, but not 51·8 GHz. The results are novel, showing distinguishing effects of low-intensity electromagnetic field of different frequencies. They could be applied in treatment of food and different products in medicine and veterinary, where E. hirae plays an important role.


Subject(s)
Enterococcus hirae/growth & development , Enterococcus hirae/radiation effects , Electromagnetic Radiation , Enterococcus hirae/chemistry , Enterococcus hirae/ultrastructure , Kinetics , Microscopy, Electron, Scanning
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