969 research outputs found
Hybrid Methods in Iron-Sulfur Cluster Biogenesis
Full text linkHybrid methods, which combine and integrate several biochemical and biophysical techniques, have rapidly caught up in the last twenty years to provide a way to obtain a fuller description of proteins and molecular complexes with sizes and complexity otherwise not easily affordable. Here, we review the use of a robust hybrid methodology based on a mixture of NMR, SAXS, site directed mutagenesis and molecular docking which we have developed to determine the structure of weakly interacting molecular complexes. We applied this technique to gain insights into the structure of complexes formed amongst proteins involved in the molecular machine, which produces the essential iron-sulfur cluster prosthetic groups. Our results were validated both by X-ray structures and by other groups who adopted the same approach. We discuss the advantages and the limitations of our methodology and propose new avenues, which could improve it
A Florentine family in crisis: the Strozzi in the fifteenth century.
No full textPhDIn 1434 the Strozzi lineage had held a leading position in
Florentine society and government for at least one hundred and fifty
years, and was one of the largest and wealthiest of the city's
patrician lineages. The records of the catasto of 1427 and of the
scrutiny of 1433 are used to give a profile of the dominant social,
economic and political position of the Strozzi before the advent of
Medicean dominance. Their record of electoral success, and the
political and cultural leadership of influential and respected men
such as Palla di Nofri and Matteo di Simone, with other factors, put
the Strozzi amongst the greatest enemies of the victorious Medicean
regime of late 1434. The effects of political opposition and exile
on the lineage are examined both directly, through records of office-holding,
and indirectly through such indicators as marriage alliances
and household wealth. The two most prominent lines of the Strozzi
were exiled after 1434. Palla di Nofri's life and preoccupations in
his Paduan exile are examined, together with the lives of his sons;
none of these Strozzi ever returned to Florence, pursued as they were
by the enmity of the Medicean regime. The very different careers of
Filippo di Matteo and his brother Lorenzo are also examined: how they
succeeded in founding a lucrative bank in Naples, and in returning to
Florence to 'rebuild' (rifare) the position of the Strozzi lineage
there. The final decades of the century saw the Strozzi in an
economically more secure position, due substantially to the efforts
of Filippo. Except for a very small number of its members admitted
into the regime, most of the lineage is here shown to have remained
excluded from significant political office until after the fall of
the Medici regime in 1494
Selective observation of the disordered import signal of a globular protein by in‐cell NMR: The example of frataxins
Full text linkAbstractWe have exploited the capability of in‐cell NMR to selectively observe flexible regions within folded proteins to carry out a comparative study of two members of the highly conserved frataxin family which are found both in prokaryotes and in eukaryotes. They all contain a globular domain which shares more than 50% identity, which in eukaryotes is preceded by an N‐terminal tail containing the mitochondrial import signal. We demonstrate that the NMR spectrum of the bacterial ortholog CyaY cannot be observed in the homologous E. coli system, although it becomes fully observable as soon as the cells are lysed. This behavior has been observed for several other compact globular proteins as seems to be the rule rather than the exception. The NMR spectrum of the yeast ortholog Yfh1 contains instead visible signals from the protein. We demonstrate that they correspond to the flexible N‐terminal tail indicating that this is flexible and unfolded. This flexibility of the N‐terminus agrees with previous studies of human frataxin, despite the extensive sequence diversity of this region in the two proteins. Interestingly, the residues that we observe in in‐cell experiments are not visible in the crystal structure of a Yfh1 mutant designed to destabilize the first helix. More importantly, our results show that, in cell, the protein is predominantly present not as an aggregate but as a monomeric species.</jats:p
MD and NMR studies of alpha-bungarotoxin surface accessibility
No full textProtein surface accessibility represents a dimension of structural biology which has not been discussed in details so far, in spite of its fundamental role in controlling the molecular recognition process. In the present report the surface accessibility of alpha-bungarotoxin, a small and well characterized protein, has been investigated by analyzing its interaction with solvent and paramagnetic molecules in an integrated way. The presence of strong hydration sites, identified by a combined analysis of MD simulation and NMR results, seems to prevent the access of Gd(III)DTPA-BMA to the protein surface. On the contrary, the limited hydration of the alpha-bungarotoxin active site favors frequent encounters between the paramagnetic probe and the protein in the latter region. All the data obtained here for alpha-bungarotoxin suggest that shape and stability of the solvation shell control its surface accessibility and, hence, intermolecular interactions in a way which could be common to many other proteins
Functionally distinct mutations within AcrB underpin antibiotic resistance in different lifestyles
Full text linkAntibiotic resistance is a pressing healthcare challenge and is mediated by various mechanisms, including the active export of drugs via multidrug efflux systems, which prevent drug accumulation within the cell. Here, we studied how Salmonella evolved resistance to two key antibiotics, cefotaxime and azithromycin, when grown planktonically or as a biofilm. Resistance to both drugs emerged in both conditions and was associated with different substitutions within the efflux-associated transporter, AcrB. Azithromycin exposure selected for an R717L substitution, while cefotaxime for Q176K. Additional mutations in ramR or envZ accumulated concurrently with the R717L or Q176K substitutions respectively, resulting in clinical resistance to the selective antibiotics and cross-resistance to other drugs. Structural, genetic, and phenotypic analysis showed the two AcrB substitutions confer their benefits in profoundly different ways. R717L reduces steric barriers associated with transit through the substrate channel 2 of AcrB. Q176K increases binding energy for cefotaxime, improving recognition in the distal binding pocket, resulting in increased efflux efficiency. Finally, we show the R717 substitution is present in isolates recovered around the world
Vues présentes sur le celtique cisalpin
No full textPresent views on Cisalpine Celtic.
During the last twenty years, the knowledge of Lepontic and of Italian Gaulish has been notably enriched by very important finds, and also by numerous studies. The author proposes a synthesis of all new linguistical and cultural data provided by epigraphy in this particular resort.Dans les vingt dernières années la connaissance du lépontique et du gaulois d’Italie a été notablement enrichie par certaines trouvailles fort importantes ainsi que par de nombreuses études. Tableau synthétique des nouvelles données linguistiques et culturelles apportées par l’épigraphie de ce domaine.Motta Filippo. Vues présentes sur le celtique cisalpin. In: Etudes Celtiques, vol. 29, 1992. Actes du IXe congrès international d'études celtiques. Paris, 7-12 juillet 1991. Deuxième partie : Linguistique, littératures. pp. 311-318
The N‐terminus of mature human frataxin is intrinsically unfolded
Full text linkFrataxin is a highly conserved nuclear‐encoded mitochondrial protein whose deficiency is the primary cause of Friedreich’s ataxia, an autosomal recessive neurodegenerative disease. The frataxin structure comprises a well‐characterized globular domain that is present in all species and is preceded in eukaryotes by a non‐conserved N‐terminal tail that contains the mitochondrial import signal. Little is known about the structure and dynamic properties of the N‐terminal tail. Here, we show that this region is flexible and intrinsically unfolded in human frataxin. It does not alter the iron‐binding or self‐aggregation properties of the globular domain. It is therefore very unlikely that this region could be important for the conserved functions of the protein.</jats:p
Bacterial frataxin CyaY is the gatekeeper of iron-sulfur cluster formation catalyzed by IscS
No full textFrataxin is an essential mitochondrial protein whose reduced expression causes Friedreich's ataxia (FRDA), a lethal neurodegenerative disease. It is believed that frataxin is an iron chaperone that participates in iron metabolism. We have tested this hypothesis using the bacterial frataxin ortholog, CyaY, and different biochemical and biophysical techniques. We observe that CyaY participates in iron-sulfur (Fe-S) cluster assembly as an iron-dependent inhibitor of cluster formation, through binding to the desulfurase IscS. The interaction with IscS involves the iron binding surface of CyaY, which is conserved throughout the frataxin family. We propose that frataxins are iron sensors that act as regulators of Fe-S cluster formation to fine-tune the quantity of Fe-S cluster formed to the concentration of the available acceptors. Our observations provide new perspectives for understanding FRDA and a mechanistic model that rationalizes the available knowledge on frataxin. © 2009 Nature America, Inc. All rights reserved
Occhi pieni e mani vaganti. Movimenti, emozioni, astrazioni.
No full textFilippo FIMIANI, Occhi pieni e mani vaganti. Movimenti, emozioni, astrazioni, «Fata Morgana», 12/2010, pp. 147-164. ISSN 1970-5786.
ITALIANO. Una piccola sequenza da un episodio della serie televisiva Mad Men in cui la relazione sensoriale emozionale tra lo spettatore e un corpo in movimento, precisamente un corpo danzante, è l’oggetto di questo articolo, che intende così verificare la cosiddetta naturalizzazione della fenomenologia e del corpo vissuto operata dalle scienze cognitive applicate agli studi visuali. L’autore mostra anche i rapporti tra la cosiddetta simulazione incarnata e una memoria simbolica e iconografica profonda, e discute in particolare la polarità tra proiezione e introiezione, attività e passività nell’esperienza empatica delle immagini in movimento.A short filmic example from the AMC's award-winning series Mad Men in which the sensorial and emotional relationship between the spectator and a moving body, i.e., a dancing body, is explicitly put into play. The essay wants to deal with the so called naturalization of phenomenology and of the living body, trained by the cognitive sciences reading the visuality. The author shows the relationship between the so called ‘embodied Simulation’ and a symbolic and iconographic deep memory of the pictures, and he gives an in-depth attention to the polarity between projections and introjections, activity and passivity of the spectator’s body during the empathic experience of the moving pictures
Dimerization of the C-type lectin-like receptor CD93 promotes its binding to Multimerin-2 in endothelial cells.
Full text linkBlocking the signaling activated by the plasma membrane receptor CD93 has recently been demonstrated a useful tool in antiangiogenic treatment and oncotherapy. In the proliferating endothelium, CD93 regulates cell adhesion, migration, and vascular maturation, yet it is unclear how CD93 interacts with the extracellular matrix activating signaling pathways involved in the vascular remodeling. Here for the first time we show that in endothelial cells CD93 is structured as a dimer and that this oligomeric form is physiologically instrumental for the binding of CD93 to its ligand Multimerin-2. Crystallographic X-ray analysis of recombinant CD93 reveals the crucial role played by the C-type lectin-like and sushi-like domains in arranging as an antiparallel dimer to achieve a functional binding state, providing key information for the future design of new drugs able to hamper CD93 function in neovascular pathologies
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