Pressure as a limiting factor for life

Facts concerning the stability and functioning of key biomolecular components suggest that cellular life should no longer be viable above a few thousand atmospheres (200–300 MPa). However, organisms are seen to survive in the laboratory to much higher pressures, extending into the GPa or even tens of GPa ranges. This is causing main questions to be posed concerning the survival mechanisms of simple to complex organisms. Understanding the ultimate pressure survival of organisms is critical for food sterilization and agricultural products conservation technologies. On Earth the deep biosphere is limited in its extent by geothermal gradients but if life forms exist in cooler habitats elsewhere then survival to greater depths must be considered. The extent of pressure resistance and survival appears to vary greatly with the timescale of the exposure. For example, shock experiments on nanosecond timescales reveal greatly enhanced survival rates extending to higher pressure. Some organisms could survive bolide impacts thus allowing successful transport between planetary bodies. We summarize some of the main questions raised by recent results and their implications for the survival of life under extreme compression conditions and its possible extent in the laboratory and throughout the universe

On the Thermal Expansion of Water and the Phase Behavior of Macromolecules in Aqueous Solution

Water is crucial for the existence of life as we know it, and many have wondered what makes water so special. Here we point out the analogies between the pressure-temperature dependence of the isobaric thermal expansion of water (α p ) and the pressure-temperature phase behavior of macromolecules in aqueous solutions. We suggest that α p could be the key to understand why water is to be the so-called 'matrix of life'

The Right-Handed Parallel β-Helix Topology of Erwinia chrysanthemi Pectin Methylesterase Is Intimately Associated with Both Sequential Folding and Resistance to High Pressure.

peer reviewedThe complex topologies of large multi-domain globular proteins make the study of their folding and assembly particularly demanding. It is often characterized by complex kinetics and undesired side reactions, such as aggregation. The structural simplicity of tandem-repeat proteins, which are characterized by the repetition of a basic structural motif and are stabilized exclusively by sequentially localized contacts, has provided opportunities for dissecting their folding landscapes. In this study, we focus on the Erwinia chrysanthemi pectin methylesterase (342 residues), an all-β pectinolytic enzyme with a right-handed parallel β-helix structure. Chemicals and pressure were chosen as denaturants and a variety of optical techniques were used in conjunction with stopped-flow equipment to investigate the folding mechanism of the enzyme at 25 °C. Under equilibrium conditions, both chemical- and pressure-induced unfolding show two-state transitions, with average conformational stability (ΔG° = 35 ± 5 kJ·mol-1) but exceptionally high resistance to pressure (Pm = 800 ± 7 MPa). Stopped-flow kinetic experiments revealed a very rapid (τ < 1 ms) hydrophobic collapse accompanied by the formation of an extended secondary structure but did not reveal stable tertiary contacts. This is followed by three distinct cooperative phases and the significant population of two intermediate species. The kinetics followed by intrinsic fluorescence shows a lag phase, strongly indicating that these intermediates are productive species on a sequential folding pathway, for which we propose a plausible model. These combined data demonstrate that even a large repeat protein can fold in a highly cooperative manner

Pressure tolerance of Artemia cysts compressed in water medium

The high pressure tolerance of cysts of Artemia salina was investigated up to several GPa in water. No survival was observed after exposure to 1.0 GPa for 15 min. After exposure to 2.0 GPa for the same time duration, the hatching rate had recovered to 33%, but decreased to 8% following compression at 7.5 GPa. This contrasts with results using Fluorinert™ as the pressure-transmitting medium where 80–88% recovery was observed. The lower survival rate in water is accompanied by swelling of the eggs, indicating that liquid H2O close to the ice-VI crystallization pressure penetrated inside the eggs. This pressure exceeds the stability limit for proteins and other key biomolecules components within the embryos that could not be resuscitated. Rehydration takes several minutes and so was not completed for all samples compressed to higher pressures, prior to ice-VI formation, resulting in renewed survival. However H2O penetration inside the shell resulted in increased mortalit

In Vivo Water Dynamics in Shewanella oneidensis Bacteria at High Pressure

Abstract: Following observations of survival of microbes and other life forms in deep subsurface environments it is necessary to understand their biological functioning under high pressure conditions. Key aspects of biochemical reactions and transport processes within cells are determined by the intracellular water dynamics. We studied water diffusion and rotational relaxation in live Shewanella oneidensis bacteria at pressures up to 500 MPa using quasi-elastic neutron scattering (QENS). The intracellular diffusion exhibits a significantly greater slowdown (by −10–30%) and an increase in rotational relaxation times (+10–40%) compared with water dynamics in the aqueous solutions used to resuspend the bacterial samples. Those results indicate both a pressure-induced viscosity increase and slowdown in ionic/macromolecular transport properties within the cells affecting the rates of metabolic and other biological processes. Our new data support emerging models for intracellular organisation with nanoscale water channels threading between macromolecular regions within a dynamically organized structure rather than a homogenous gel-like cytoplasm

Generic Mechanism of Emergence of Amyloid Protofilaments from Disordered Oligomeric aggregates

The presence of oligomeric aggregates, which is often observed during the process of amyloid formation, has recently attracted much attention since it has been associated with neurodegenerative conditions such as Alzheimer's and Parkinson's diseases. We provide a description of a sequence-indepedent mechanism by which polypeptide chains aggregate by forming metastable oligomeric intermediate states prior to converting into fibrillar structures. Our results illustrate how the formation of ordered arrays of hydrogen bonds drives the formation of beta-sheets within the disordered oligomeric aggregates that form early under the effect of hydrophobic forces. Initially individual beta-sheets form with random orientations, which subsequently tend to align into protofilaments as their lengths increases. Our results suggest that amyloid aggregation represents an example of the Ostwald step rule of first order phase transitions by showing that ordered cross-beta structures emerge preferentially from disordered compact dynamical intermediate assemblies.Comment: 14 pages, 4 figure

Comparative Fourier transform infrared spectroscopy study of cold-, pressure-, and heat-induced unfolding and aggregation of myoglobin.

We studied the cold unfolding of myoglobin with Fourier transform infrared spectroscopy and compared it with pressure and heat unfolding. Because protein aggregation is a phenomenon with medical as well as biotechnological implications, we were interested in both the structural changes as well as the aggregation behavior of the respective unfolded states. The cold- and pressure-induced unfolding both yield a partially unfolded state characterized by a persistent amount of secondary structure, in which a stable core of G and H helices is preserved. In this respect the cold- and pressure-unfolded states show a resemblance with an early folding intermediate of myoglobin. In contrast, the heat unfolding results in the formation of the infrared bands typical of intermolecular antiparallel beta-sheet aggregation. This implies a transformation of alpha-helix into intermolecular beta-sheet. H/2H-exchange data suggest that the helices are first unfolded and then form intermolecular beta-sheets. The pressure and cold unfolded states do not give rise to the intermolecular aggregation bands that are typical for the infrared spectra of many heat-unfolded proteins. This suggests that the pathways of the cold and pressure unfolding are substantially different from that of the heat unfolding. After return to ambient conditions the cold- or pressure-treated proteins adopt a partially refolded conformation. This aggregates at a lower temperature (32 degrees C) than the native state (74 degrees C)

Infrared spectroscopy and ultrasonic velocimetry as a tool for probing conformational flexibility of proteins

Because of the crucial importance of structural fluctuations for function and stability of proteins, there is a strong interest for the relationships between structural fluctuations, the parameters of protein denaturation and the kinetics of H/D-exchange. Structural fluctuations can be described by volume and entropy fluctuations and these quantities are accessible via the isothermal compressibility, the thermal expansion and the isobaric heat capacity.status: publishe