The Met80Ala and Tyr67His/Met80Ala mutants of human cytochrome cshed light on the reciprocal role of Met80 and Tyr67 in regulating ligand access into the heme pocket.

The spectroscopic and functional properties of the single Met80Ala and double Tyr67His/Met80Ala mutants of human cytochrome c have been investigated in their ferric and ferrous forms, and in the presence of different ligands, in order to clarify the reciprocal effect of these two residues in regulating the access of exogenous molecules into the heme pocket. In the ferric state, both mutants display an aquo high spin and a low spin species. The latter corresponds to an OH- ligand in Met80Ala but to a His in the double mutant. The existence of these two species is also reflected in the functional behavior of the mutants. The observation that (i) a significant peroxidase activity is present in the Met80Ala mutants, (ii) the substitution of the Tyr67 by His leads to only a slight increase of the peroxidase activity in the Tyr67His/Met80Ala double mutant with respect to wild type, while the Tyr67His mutant behaves as wild type, as previously reported, suggests that the peroxidase activity of cytochrome c is linked to an overall conformational change of the heme pocket and not only to the disappearance of the Fe-Met80 bond. Therefore, in human cytochrome c there is an interplay between the two residues at positions 67 and 80 that affects the conformation of the distal side of the heme pocket, and thus the sixth coordination of the hem

Evaluation of Seismic Vulnerability of Santa Maria del Mar in Barcelona by an Integrated Approach Based on Terrestrial Laser Scanner and Finite Element Modelling

Structural assessment of historical constructions is a particular issue as these structures were built with technical knowledge and construction methods based on in-field experience and on the observation of previous failures and successes of similar buildings. Their structural behavior is influenced by numerous aspects, sometimes not completely known (geometry, structural schemes and details, material mechanical properties), which often results in historical structures inadequate to achieve the safety level required for new constructions. In the case of earthquakes, these structures can suffer heavy local or global damages that sometimes lead to collapse. Thus static and seismic vulnerability assessment of historical buildings should therefore be performed by proper methodologies that differ from those used for new constructions. In Italy, an official document (Direttiva del Presidente del Consiglio dei Ministri [DPCM] 2007, revised in 2011) was released, defining a step-by-step procedure for data acquisition and vulnerability assessment of Cultural Heritage buildings. Such procedures can be coupled with new survey technologies such as terrestrial laser scanning (TLS), which, for performing a high-detailed three-dimensional geometrical survey, can provide detailed structural models refining the assessment results. In this study, the DPCM procedure is applied to Santa Maria del Mar Church in Barcelona (Spain) for seismic vulnerability assessment, using TLS three-dimensional survey and finite element analysis

High-Lift Devices Topology Optimisation using Structured-Chromosome Genetic Algorithm

This paper addresses the problem of including the choice of the High-Lift Devices (HLDs) configuration as a decision variable of an automatic optimisation tool. This task requires the coupling of an estimation routine and an optimisation algorithm. For the former, SU2 flow solver has been used. The Structured-Chromosome Genetic Algorithm (SCGA) optimiser has been employed to search for the optimal HLD. SCGA can overcome the limitations dictated by standard fixed-size continuous optimisation algorithms. Indeed, using hierarchical formulations, it can manage configurational decisions that are conventionally the responsibility of expert designers. The search algorithm bases its strategy on revised genetic operators conceived for handling hierarchical search spaces. The presented research not only shows the practicability of delegating to a specialised optimisation algorithm the complete HLD design but is intended to be a proof of concept for the whole field of multidisciplinary design optimisation. Indeed, the aerospace sector as a whole would benefit by reducing human intervention from the decision process

Unravelling the Non-Native Low-Spin State of the Cytochrome c-Cardiolipin Complex: Evidence of the Formation of a His-Ligated Species Only

The interaction between cytochrome c (Cyt c) and cardiolipin (CL) plays a vital role in the early stages of apoptosis. The binding of CL to Cyt c induces a considerable increase in its peroxidase activity that has been attributed to the partial unfolding of the protein, dissociation of the Met80 axial ligand, and formation of non-native conformers. Although the interaction between Cyt c and CL has been extensively studied, there is still no consensus regarding the conformational rearrangements of Cyt c that follow the protein-lipid interaction. To rationalize the different results and gain better insight into the Cyt c-CL interaction, we have studied the formation of the CL complex of the horse heart wild-type protein and selected mutants in which residues considered to play a key role in the interaction with CL (His26, His33, Lys72, Lys73, and Lys79) have been mutated. The analysis was conducted at both room temperature and low temperatures via ultraviolet-visible absorption, resonance Raman, and electron paramagnetic resonance spectroscopies. The trigger and the sequence of CL-induced structural variations are discussed in terms of disruption of the His26-Pro44 hydrogen bond. We unequivocally identify the sixth ligand in the partially unfolded, non-native low-spin state that Cyt c can adopt following the protein-lipid interaction, as a His ligation, ruling out the previously proposed involvement of a Lys residue or an OH- ion