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Denaturation Process (denaturation + process)
Selected AbstractsDirect detection of the protein quaternary structure and denatured entity by small-angle scattering: guanidine hydrochloride denaturation of chaperonin protein GroELJOURNAL OF APPLIED CRYSTALLOGRAPHY, Issue 1 2002Yasutaka Seki A change in the higher-order structure of an oligomeric protein is directly detectable by small-angle scattering. A small-angle X-ray scattering (SAXS) study of the denaturation process of the chaperonin protein GroEL by guanidine hydrochloride (GdnHCl) showed that the disappearance of the quaternary structure can be monitored by using a Kratky plot of the scattered intensities, demonstrating the advantage of the SAXS method over other indirect methods, such as light scattering, circular dichroism (CD), fluorescence and sedimentation. The collapse of the quaternary structure was detected at a GdnHCl concentration of 0.8,M for a solution containing ADP (adenosine diphosphate)/Mg2+(2,mM)/K+. From pairwise plots of the change in forward scattering intensity J(0)/C (weight-average molecular weight) and the z -average (root mean square) radius of gyration against the GdnHCl concentration, the stability and nature of the denatured protein can be determined. The SAXS results suggest that the GroEL tetradecamer directly dissociates to the unfolded coil without going through a globular monomer state. The denatured ensemble is not a single unfolded monomer coil particle, but some mixture of entangled aggregates and a monomer of the coil molecules. Small-angle scattering is a powerful method for the detection and study of changes in quaternary and higher-order structures of oligomeric proteins. [source] Monitoring of unfolding of metallo-proteins by electrospray ionization mass spectrometryJOURNAL OF MASS SPECTROMETRY (INCORP BIOLOGICAL MASS SPECTROMETRY), Issue 5 2003Vincenzo Cunsolo Abstract An electrospray ionisation (ESI) mass spectrometric method for the determination of the equilibrium constant and free energy (,G) of protein unfolding was used to monitor the denaturation process at different pH of three metallo-proteins, i.e. wild-type copper azurin, zinc azurin and wild-type amicyanin. The time course of the unfolding process was followed by dissolving the proteins under denaturing conditions (methanol,water (1 : 1, v/v)) at different pH (2.5, 3.0, 3.5) and recording ESI spectra at time intervals. The spectra showed two series of peaks, corresponding to the native holo-protein and the unfolded apo-protein. From the intensity ratio of these two series of peaks at increasing time and at equilibrium, the equilibrium constants for the unfolding process for the three proteins could be determined. From these equilibrium constants a ,G° derivation was attempted. The ,G° values obtained decrease with decrease in pH, in agreement with the expected reduction of conformational stability of proteins at lower pH. The results obtained confirm that ESI-MS can be used for monitoring of unfolding process and to derive quantitative thermodynamic data. Copyright © 2003 John Wiley & Sons, Ltd. [source] Spectroscopic study of the physical properties making trehalose a stabilizing and shelf life extending compound in food industryQUALITY ASSURANCE & SAFETY OF CROPS & FOOD, Issue 2 2010S. Magazł Abstract Introduction Trehalose, a glass-forming bioprotectant disaccharide, has been demonstrated to possess significant potential within the food industry. It does not interact with reactive molecules such as amino groups from peptides and proteins, preventing the degradation and aggregation due to Maillard reactions. Objective This paper aims to review at the molecular level the effects of trehalose on the structural and dynamical properties of water and on protein to highlight the stabilization and conservation properties on food products. Results and Conclusions The experimental findings presented show that water molecules are arranged in presence of trehalose in a particular configuration which avoids ice formation, so limiting damage due to freezing and cooling. On the other hand, homologous disaccharides, and trehalose to a greater extent, slow down the dynamics of water with a significant influence on the biological activity. These results imply that trehalose has a greater ability to bind volatile substances and deliver superior bioprotective effectiveness. Furthermore trehalose is shown to be incapable of taking part in the denaturation process of lysozyme under thermal stress. [source] High thermal and chemical stability of Thermus thermophilus seven-iron ferredoxinFEBS JOURNAL, Issue 23 2003Linear clusters form at high pH on polypeptide unfolding To probe the stability of the seven-iron ferredoxin from Thermus thermophilus (FdTt), we investigated its chemical and thermal denaturation processes in solution. As predicted from the crystal structure, FdTt is extremely resistant to perturbation. The guanidine hydrochloride-induced unfolding transition shows a midpoint at 6.5 m (pH 7, 20 °C), and the thermal midpoint is above boiling, at 114 °C. The stability of FdTt is much lower at acidic pH, suggesting that electrostatic interactions are important for the high stability at higher pH. On FdTt unfolding at alkaline pH, new absorption bands at 520 nm and 610 nm appear transiently, resulting from rearrangement of the cubic clusters into linear three-iron species. A range of iron,sulfur proteins has been found to accommodate these novel clusters in vitro, although no biological function has yet been assigned. [source] | ||||||||||