The rod-shaped Gram-negative bacterium Escherichia coli multiplies by elongation followed binary fission. Longitudinal growth of the cell envelope and synthesis new poles are organized two protein complexes called elongasome divisome, respectively. We have analyzed spatio-temporal localization patterns many these morphogenetic proteins immunolabeling wild type strain MC4100 grown to steady state in minimal glucose medium at 28°C. This allowed direct comparison as a function age imaged phase...

Significance The shape of biological membranes is constantly remodeled and maintained out equilibrium by active proteins. functional capacity membrane deformation mainly determined the mechanical interplay between protein activity bending elasticity. In our experiments, we find that ATP synthase, a rotating synthesizes biochemical energy in cells through proton-pumping across membrane, promotes localized nonequilibrium fluctuations when reconstituted giant lipid vesicles. large deformations...

Abstract ATP, the molecule used by living organisms to supply energy many different metabolic processes, is synthesized mostly ATPase synthase using a proton or sodium gradient generated across lipid membrane. We present evidence that modified electrode surface integrating NiFeSe hydrogenase and F 1 0 ‐ATPase in membrane can couple electrochemical oxidation of H 2 synthesis ATP. This electrode‐assisted conversion gas into ATP could serve generate this biochemical fuel locally when required...

Fluorescent proteins are extraordinary tools for biology studies due to their versatility; they used extensively improve comprehension of plant-microbe interactions. The viral infection process can easily be tracked and imaged in a plant with fluorescent protein-tagged viruses. In plants, protein genes among the most commonly reporters transient RNA silencing heterologous expression assays. Fluorescence intensity is quantify accumulation by image analysis or spectroscopy extracts; however,...

Abstract Background A major bottleneck in drug delivery is the breakdown and degradation of system through endosomal/lysosomal network host cell, hampering correct interest. In nature, bacterial pathogen Listeria monocytogenes has developed a strategy to secrete Listeriolysin O (LLO) toxin as tool escape eukaryotic lysosomal upon infection, allowing it grow proliferate unharmed inside cell. Results As “proof concept”, we present here use purified His-LLO H311A mutant protein its conjugation...

Septation in Escherichia coli involves complex molecular mechanisms that contribute to the accuracy of bacterial division. The proto-ring, a made up by FtsZ, FtsA and ZipA proteins, forms at beginning process directs assembly full divisome. Central this is FtsZ protein, GTPase able assemble into ring-like structure responds several modulatory inputs including position septum midcell. connection with cell wall synthesising machinery stabilizes constriction cytoplasmic membrane. Although...

We describe the placement of cytoplasmic FtsZ protein, an essential component division septum, in nucleoid-free Escherichia coli maxicells. The absence nucleoid is accompanied maxicells by degradation SlmA protein. This together with nucleoid, prevents septum regions occupied chromosome a mechanism called occlusion (NO). A second mechanism, MinCDE system (Min) involving pole-to-pole oscillation three proteins, nonetheless remains active Both Min and NO act on polymerization FtsZ, preventing...

Summary We used bimolecular fluorescence complementation ( BiFC ) assays to detect protein‐protein interactions of all possible pairs the essential E scherichia coli proto‐ring components, FtsZ , FtsA and ZipA as well non‐essential ‐associated proteins ZapA ZapB . found an unexpected interaction between at potential cell division sites, when co‐overproduced, they induced long narrow constrictions sites that were dependent on These also uncovered was mediated by with showed in addition their...

The mitochondrion is an essential organelle enclosed by two membranes whose functionalities depend on their very specific protein and lipid compositions. Proteins from the outer mitochondrial membrane (OMM) are specialized in dynamics mitophagy, whereas proteins of inner (IMM) have dedicated functions cellular respiration apoptosis. As for lipids, OMM enriched glycerophosphatidyl choline but cardiolipin exclusively found within IMM. Though topology distribution IMM known since more than four...

The design of efficient drug delivery vectors requires versatile formulations able to simultaneously direct a multitude molecular targets and bypass the endosomal recycling pathway cells. Liposomal-based need decoration lipid surface with specific peptides fulfill functional requirements. unspecific binding is often accompanied uncontrolled thus preventing mechanisms successful therapy.We present simple synthesis anchor cysteine-terminal thiol-reactive lipids for adequate quantitative...

SecYEG translocase mediates the transport of preproteins across inner membrane Escherichia coli. SecA binds membrane-embedded protein-conducting channel with high affinity and then drives stepwise translocation through multiple cycles ATP binding hydrolysis. We have investigated kinetics nucleotide to while associated complex. Lipid-bound was separated from Se-cYEG-bound by sedimentation proteoliposomes a glycerol cushion, which maintains native state effectively removes lipid-bound...

Mitochondria are essential for the production and maintenance of ATP in eukaryotic cell. To image monitor intracellular level without cell breakage, biological chemical sensors were developed last years. Here, we have internalized a rhodamine-based sensor RSL+ into living cells monitored mitochondrial levels cultured mouse embryonic fibroblasts. evaluate robustness imaged changes under non-physiological conditions upon incubation with FCCP, oligomycin, azide, deoxyglucose or...

Adenosine-5-triphosphate (ATP) is the main energy vector in biological systems, thus its regeneration an important issue for application of many enzymes interest biocatalysis and synthetic biology. We have developed electroenzymatic ATP system consisting a gold electrode modified with floating phospholipid bilayer that allows coupling catalytic activity two membrane-bound enzymes: NiFeSe hydrogenase from Desulfovibrio vulgaris F1Fo-ATP synthase Escherichia coli. Thus, H2 used as fuel...

The SecYEG complex is a membrane-embedded channel that permits the passage of precursor proteins (preproteins) across inner membrane Escherichia coli. SecA molecular motor associates with pore and drives stepwise translocation preproteins through multiple cycles ATP binding hydrolysis. We have investigated conformational state soluble SecYEG-bound using single tryptophan mutants SecA. fluorescence spectral properties tryptophans their accessibility to quencher acrylamide demonstrate...

Cancer cell mitochondria represent an attractive target for oncological treatment as they have unique hallmarks that differ from their healthy counterparts, the presence of a stronger membrane potential can be exploited to specifically accumulate cytotoxic cationic molecules. Here, we explore selective effect 10-N-nonyl acridine orange (NAO) on human lung carcinoma H520 cells and compare them with primary fibroblasts. NAO is lipophilic positively charged molecule promotes mitochondrial...

Abstract ATP, the molecule used by living organisms to supply energy many different metabolic processes, is synthesized mostly ATPase synthase using a proton or sodium gradient generated across lipid membrane. We present evidence that modified electrode surface integrating NiFeSe hydrogenase and F 1 0 ‐ATPase in membrane can couple electrochemical oxidation of H 2 synthesis ATP. This electrode‐assisted conversion gas into ATP could serve generate this biochemical fuel locally when required...