54 research outputs found
Mechanism of Protein Kinetic Stabilization by Engineered Disulfide Crosslinks
The impact of disulfide bonds on protein stability goes beyond simple equilibrium thermodynamics effects associated with the conformational entropy of the unfolded state. Indeed, disulfide crosslinks may play a role in the prevention of dysfunctional association and strongly affect the rates of irreversible enzyme inactivation, highly relevant in biotechnological applications. While these kinetic-stability effects remain poorly understood, by analogy with proposed mechanisms for processes of protein aggregation and fibrillogenesis, we propose that they may be determined by the properties of sparsely-populated, partially-unfolded intermediates. Here we report the successful design, on the basis of high temperature molecular-dynamics simulations, of six thermodynamically and kinetically stabilized variants of phytase from Citrobacter braakii (a biotechnologically important enzyme) with one, two or three engineered disulfides. Activity measurements and 3D crystal structure determination demonstrate that the engineered crosslinks do not cause dramatic alterations in the native structure. The inactivation kinetics for all the variants displays a strongly non-Arrhenius temperature dependence, with the time-scale for the irreversible denaturation process reaching a minimum at a given temperature within the range of the denaturation transition. We show this striking feature to be a signature of a key role played by a partially unfolded, intermediate state/ensemble. Energetic and mutational analyses confirm that the intermediate is highly unfolded (akin to a proposed critical intermediate in the misfolding of the prion protein), a result that explains the observed kinetic stabilization. Our results provide a rationale for the kinetic-stability consequences of disulfide-crosslink engineering and an experimental methodology to arrive at energetic/structural descriptions of the sparsely populated and elusive intermediates that play key roles in irreversible protein denaturation.This work was supported by grants BIO2009-09562, CSD2009-00088 from the Spanish Ministry of Science and Innovation, and FEDER Funds (JMS-R)
Gravitational-wave research as an emerging field in the Max Planck Society. The long roots of GEO600 and of the Albert Einstein Institute
On the occasion of the 50th anniversary since the beginning of the search for
gravitational waves at the Max Planck Society, and in coincidence with the 25th
anniversary of the foundation of the Albert Einstein Institute, we explore the
interplay between the renaissance of general relativity and the advent of
relativistic astrophysics following the German early involvement in
gravitational-wave research, to the point when gravitational-wave detection
became established by the appearance of full-scale detectors and international
collaborations. On the background of the spectacular astrophysical discoveries
of the 1960s and the growing role of relativistic astrophysics, Ludwig Biermann
and his collaborators at the Max Planck Institute for Astrophysics in Munich
became deeply involved in research related to such new horizons. At the end of
the 1960s, Joseph Weber's announcements claiming detection of gravitational
waves sparked the decisive entry of this group into the field, in parallel with
the appointment of the renowned relativist Juergen Ehlers. The Munich area
group of Max Planck institutes provided the fertile ground for acquiring a
leading position in the 1970s, facilitating the experimental transition from
resonant bars towards laser interferometry and its innovation at increasingly
large scales, eventually moving to a dedicated site in Hannover in the early
1990s. The Hannover group emphasized perfecting experimental systems at pilot
scales, and never developed a full-sized detector, rather joining the LIGO
Scientific Collaboration at the end of the century. In parallel, the Max Planck
Institute for Gravitational Physics (Albert Einstein Institute) had been
founded in Potsdam, and both sites, in Hannover and Potsdam, became a unified
entity in the early 2000s and were central contributors to the first detection
of gravitational waves in 2015.Comment: 94 pages. Enlarged version including new results from further
archival research. A previous version appears as a chapter in the volume The
Renaissance of General Relativity in Context, edited by A. Blum, R. Lalli and
J. Renn (Boston: Birkhauser, 2020
Religiosity, spirituality in relation to disordered eating and body image concerns: A systematic review
Short-Term Reservoir Optimization for Flood Mitigation under Meteorological and Hydrological Forecast Uncertainty
Onyx embolization of a thoracolumbar perimedullary spinal arteriovenous fistula in an infant presenting with subarachnoid and intraventricular hemorrhage
An efficient method for stochastic optimal control with joint chance constraints for nonlinear systems
- …