Alpha-helical destabilization of the Bcl-2-BH4-domain peptide abolishes its ability to inhibit the IP3 receptor

The anti-apoptotic Bcl-2 protein is the founding member and namesake of the Bcl-2-protein family. It has recently been demonstrated that Bcl-2, apart from its anti-apoptotic role at mitochondrial membranes, can also directly interact with the inositol 1,4,5-trisphosphate receptor (IP3R), the primary Ca2+-release channel in the endoplasmic reticulum (ER). Bcl-2 can thereby reduce pro-apoptotic IP3R-mediated Ca2+ release from the ER. Moreover, the Bcl-2 homology domain 4 (Bcl-2-BH4) has been identified as essential and sufficient for this IP3R-mediated anti-apoptotic activity. In the present study, we investigated whether the reported inhibitory effect of a Bcl-2-BH4 peptide on the IP (3)R1 was related to the distinctive alpha-helical conformation of the BH4 domain peptide. We therefore designed a peptide with two glycine "hinges" replacing residues I14 and V15, of the wild-type Bcl-2-BH4 domain (Bcl-2-BH4-IV/GG). By comparing the structural and functional properties of the Bcl-2-BH4-IV/GG peptide with its native counterpart, we found that the variant contained reduced alpha-helicity, neither bound nor inhibited the IP (3)R1 channel, and in turn lost its anti-apoptotic effect. Similar results were obtained with other substitutions in Bcl-2-BH4 that destabilized the alpha-helix with concomitant loss of IP3R inhibition. These results provide new insights for the further development of Bcl-2-BH4-derived peptides as specific inhibitors of the IP3R with significant pharmacological implications

Stress induced Salmonella Typhimurium re-excretion by pigs is associated with cortisol induced increased intracellular proliferation in porcine macrophages

Infections of pigs with Salmonella enterica subspecies enterica serovar Typhimurium (Salmonella Typhimurium) often result in the development of carriers that intermittently excrete Salmonella in very low numbers. During periods of stress, recrudescence of Salmonella may occur

Self-focusing in processes of laser generation of highly-charged and high-energy heavy ions

Laser-beam interaction with expanding plasma was investigated using the PALS high-power iodine-laser system. The interaction conditions are significantly changing with the laser focus spot position. The decisive role of the laser-beam self-focusing, participating in the production of ions with the highest charge states, was proved

Study of shock waves generation, hot electron production and role of parametric instabilities in an intensity regime relevant for the shock ignition

We present experimental results at intensities relevant to Shock Ignition obtained at the sub-ns Prague Asterix Laser System in 2012 . We studied shock waves produced by laser-matter interaction in presence of a pre-plasma. We used a first beam at 1ω (1315 nm) at 7 × 10 13 W/cm 2 to create a pre-plasma on the front side of the target and a second at 3ω (438 nm) at ∼ 10 16 W/cm 2 to create the shock wave. Multilayer targets composed of 25 (or 40 μm) of plastic (doped with Cl), 5 μm of Cu (for Kα diagnostics) and 20 μm of Al for shock measurement were used. We used X-ray spectroscopy of Cl to evaluate the plasma temperature, Kα imaging and spectroscopy to evaluate spatial and spectral properties of the fast electrons and a streak camera for shock breakout measurements. Parametric instabilities (Stimulated Raman Scattering, Stimulated Brillouin Scattering and Two Plasmon Decay) were studied by collecting the back scattered light and analysing its spectrum. Back scattered energy was measured with calorimeters. To evaluate the maximum pressure reached in our experiment we performed hydro simulations with CHIC and DUED codes. The maximum shock pressure generated in our experiment at the front side of the target during laser-interaction is 90 Mbar. The conversion efficiency into hot electrons was estimated to be of the order of ∼ 0.1% and their mean energy in the order ∼50 keV. Content from this work may be used under the terms of the Creative Commons Attribution 3.0 licence. Any further distributio

Angular distributions of ions emitted from laser plasma produced at various irradiation angles and laser intensities

AbstractAngular distributions of currents and velocities (energies) of ions produced at various target irradiation angles and laser intensities ranged from 1010 W/cm2 to 1017 W/cm2 were analyzed. It was confirmed that for low laser intensities the ion current distributions are always peaked along the target normal. However, at laser intensities comparable to or higher than 1014 W/cm2, the preferred direction of ion emission strongly depends on the irradiation geometry (laser focus setting, the irradiation angle), and can be off the target normal. This is very likely caused by the non-linear interaction of the laser beam with produced plasma, in particular, by the action of ponderomotive forces and the laser beam self-focusing

Factors influencing parameters of laser ion sources

Various applications demand various kinds of ions. Charge state, energy and the amount of laser produced ions depend, primary, on the wavelength, the energy, the pulse duration, and the focusing ability of the laser used. Angle of the target irradiation, angle of the ion extraction (recording), and mainly the focus setting may significantly influence especially the portion of ions with the highest charge states. The participation of non-linear processes on the generation of ions with extremely high parameters is demonstrated. The observed effects support the idea of a longitudinal structure of the self-focused laser beam with a space period of ∼200 µm

Multiply charged ions from iodine laser-produced plasma of medium- and high-Z targets

Maximum charge states of ions registered in the far expansion zone from laser-produced plasma of Al, Co, Ni, Cu, Ta, W, Pt, Au, Pb, and Bi are presented. The Thomson parabola spectrometer was used to display a general view of the ion species of an expanding plasma while detailed ion charge-energy spectra were determined by the cylindrical electrostatic ion energy analyzer. The current densities of highly charged ion groups above 20 mA/cm2 were measured by use of an ion collector at a distance of 1 m from the target. The photodissociation iodine laser system PERUN (λ = 1.315 μm, power density up to 1015 W cm−2) was employed as a drive

Sphingosine kinase 1 overexpression induces MFN2 fragmentation and alters mitochondrial matrix Ca2+ handling in HeLa cells

Sphingosine kinase 1 (SKI) converts sphingosine to the bioactive lipid sphingosine 1-phosphate (SIP). SW binds to G-protein-coupled receptors (S1PR(1-5)) to regulate cellular events, including Ca2+ signaling. The SK1/S1P axis and Ca2+ signaling both play important roles in health and disease. In this respect, Ca2+ microdomains at the mitochondria-associated endoplasmic reticulum (ER) membranes (MAMs) are of importance in oncogenesis. Mitofusin 2 (MFN2) modulates ER-mitochondria contacts, and dysregulation of MFN2 is associated with malignancies. We show that overexpression of SKI augments agonist-induced Ca2+ release from the ER resulting in increased mitochondria] matrix Ca2+. Also, overexpression of SK1 induces MFN2 fragmentation, likely through increased calpain activity. Further, expressing putative calpain-cleaved MFN2 N- and C-terminal fragments increases mitochondrial matrix Ca2+ during agonist stimulation, mimicking the SK1 overexpression in cells. Moreover, SK1 overexpression enhances cellular respiration and cell migration. Thus, SK1 regulates MFN2 fragmentation resulting in increased mitochondrial Ca2+ and downstream cellular effects.Peer reviewe

The influence of an intense laser beam interaction with preformed plasma on the characteristics of emitted ion streams

AbstractIntense laser-beam interactions with preformed plasma, preceding the laser-target interactions, significantly influence both the ion and X-ray generation. It is due to the laser pulse (its total length, the shape of the front edge, its background, the contrast, the radial homogeneity) as well as plasma (density, temperature) properties. Generation of the super fast (FF) ion groups is connected with a presence of non-linear processes. Saturated maximum of the charge states (independently on the laser intensity) is ascribed to the constant limit radius of the self-focused laser beam. Its longitudinal structure is considered as a possible explanation for the course of some experimental dependencies obtained

Ion production by lasers using high-power densities in a near infrared region

Results are presented of experiments on ion production from Ta targets using a short pulse (350-600 ps in focus) illumination with focal power densities exceeding 1014 Wcm-2 at the wavelength of an iodine photodissociation laser (1.315 μm) and its harmonics. Strong evidence of the existence of tantalum ions with the charge state +45 near the target surface was obtained by X-ray spectroscopy methods. The particle diagnostics point to the existence of frozen high charge states (4 MeV) for the highest observed charge states. A tentative theoretical explanation of the observed anomalous charge state freezing phenomenon in the expanding plasma produced by a subnanosecond laser pulse is give