From Conformation to interaction: techniques to explore the Hsp70/hsp90 network

Proteins participate in almost every cell physiological function, and to do so, they need to reach a state that allows its function by folding and/or exposing surfaces of interactions. Spontaneous folding in the cell is in general hindered by its crowded and viscous environment, which favors misfolding and nonspecific and deleterious self-interactions. To overcome this, cells have a system, in which Hsp70 and Hsp90 play a central role to aid protein folding and avoid misfolding. The topics of this review include the biophysical tools used for monitoring protein-ligand and protein-protein interactions and also some important results related to the study of molecular chaperones and heat shock proteins (Hsp), with a focus on the Hsp70/Hsp90 network. The biophysical tools and their use to probe the conformation and interaction of Hsp70 and Hsp90 are briefly reviewed168735753CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPES

A Simple Chemical Method for Rendering Wild-Type Yeast Permeable to Brefeldin A That Does Not Require the Presence of an erg6 Mutation

The present work aims to develop a growth medium to render a wild-type strain of Saccharomyces cerevisiae permeable to the antifungal drug Brefeldin A. In the current study, a synthetic medium containing 0.1% L-proline and supplemented with 3.0 × 10(-3)% SDS is employed. When Brefeldin A is added to this medium, a wild-type strain shows increased growth sensitivity and a diminished transport of the amino acid L-leucine. Since Brefeldin A exerts its effect on the endoplasmic reticulum and the Golgi apparatus, the medium permits the study of the drug effect on the intracellular traffic of L-leucine permeases

The LISA PathFinder DMU and Radiation Monitor

The LISA PathFinder DMU (Data Management Unit) flight model was formally accepted by ESA and ASD on 11 February 2010, after all hardware and software tests had been successfully completed. The diagnostics items are scheduled to be delivered by the end of 2010. In this paper we review the requirements and performance of this instrumentation, specially focusing on the Radiation Monitor and the DMU, as well as the status of their programmed use during mission operations, on which work is ongoing at the time of writing.Comment: 11 pages, 7 figures, prepared for the Proceedings of the 8th International LISA Symposium, Classical and Quantum Gravit

DIBMA nanodiscs keep α-synuclein folded

α-Synuclein (αsyn) is a cytosolic intrinsically disordered protein (IDP) known to fold into an α-helical structure when binding to membrane lipids, decreasing protein aggregation. Model membrane enable elucidation of factors critically affecting protein folding/aggregation, mostly using either small unilamellar vesicles (SUVs) or nanodiscs surrounded by membrane scaffold proteins (MSPs). Yet SUVs are mechanically strained, while MSP nanodiscs are expensive. To test the impact of lipid particle size on α-syn structuring, while overcoming the limitations associated with the lipid particles used so far, we compared the effects of large unilamellar vesicles (LUVs) and lipid-bilayer nanodiscs encapsulated by diisobutylene/maleic acid copolymer (DIBMA) on αsyn secondary-structure formation, using human-, elephant- and whale -αsyn. Our results confirm that negatively charged lipids induce αsyn folding in h-αsyn and e-αsyn but not in w-αsyn. When a mixture of zwitterionic and negatively charged lipids was used, no increase in the secondary structure was detected at 45 °C. Further, our results show that DIBMA/lipid particles (DIBMALPs) are highly suitable nanoscale membrane mimics for studying αsyn secondary-structure formation and aggregation, as folding was essentially independent of the lipid/protein ratio, in contrast with what we observed for LUVs having the same lipid compositions. This study reveals a new and promising application of polymer-encapsulated lipid-bilayer nanodiscs, due to their excellent efficiency in structuring disordered proteins such as αsyn into nontoxic α-helical structures. This will contribute to the unravelling and modelling aspects concerning protein-lipid interactions and α-helix formation by αsyn, paramount to the proposal of new methods to avoid protein aggregation and disease.info:eu-repo/semantics/publishedVersio

Identification of Regions Involved in Substrate Binding and Dimer Stabilization within the Central Domains of Yeast Hsp40 Sis1

Protein folding, refolding and degradation are essential for cellular life and are regulated by protein homeostatic processes such those that involve the molecular chaperone DnaK/Hsp70 and its co-chaperone DnaJ. Hsp70 action is initiated when proteins from the DnaJ family bind an unfolded protein for delivery purposes. In eukaryotes, the DnaJ family can be divided into two main groups, Type I and Type II, represented by yeast cytosolic Ydj1 and Sis1, respectively. Although sharing some unique features both members of the DnaJ family, Ydj1 and Sis1 are structurally and functionally distinct as deemed by previous studies, including the observation that their central domains carry the structural and functional information even in switched chimeras. In this study, we combined several biophysical tools for evaluating the stability of Sis1 and mutants that had the central domains (named Gly/Met rich domain and C-terminal Domain I) deleted or switched to those of Ydj1 to gain insight into the role of these regions in the structure and function of Sis1. The mutants retained some functions similar to full length wild-type Sis1, however they were defective in others. We found that: 1) Sis1 unfolds in at least two steps as follows: folded dimer to partially folded monomer and then to an unfolded monomer. 2) The Gly/Met rich domain had intrinsically disordered characteristics and its deletion had no effect on the conformational stability of the protein. 3) The deletion of the C-terminal Domain I perturbed the stability of the dimer. 4) Exchanging the central domains perturbed the conformational stability of the protein. Altogether, our results suggest the existence of two similar subdomains in the C-terminal domain of DnaJ that could be important for stabilizing each other in order to maintain a folded substrate-binding site as well as the dimeric state of the protein.Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Ministerio da Ciencia e Tecnologia/Conselho Nacional de Pesquisa e Desenvolvimento (MCT/CNPq

Best standards for data collection and reporting requirements on FOBs: towards a science-based FOB fishery management.

A major concern for tropical tunas, on these last years, has been the worldwide increasing use of drifting FOBs by purse seiners, which are equipped with satellite buoys and echo-sounders. The use of these floating objects has contributed to increase the catch of skipjack tuna, but also of juveniles of yellowfin and bigeye tunas. Moreover, it has increased the amount of by-catch (including some species classified as vulnerable or endangered) and has likely resulted in adverse effects on the ecology of fish and on vulnerable areas (e.g. beaching events on coral reef areas). Despite the increasing FOB use and concerns, little information is available on FOB use worldwide for an appropriate monitoring and management. Thus, FOB monitoring has become a priority in all tuna t-RFMOs. However, the data collection and reporting requirements around FOBs are not standardized and there are significant data gaps. The aim of this document is to review current requirements and procedures in place and propose standards for data collection and submission on FOBs to tRFMOs. The proposals included in this document are the result of a collaborative work between scientists and the fishing industry

Forest Fires, Land Use Changes and Their Impact on Hydrological Balance in Temperate Forests of Central Mexico

Temperate forests play a fundamental role in the provision, regulation, and support of hydrological environmental services, but they are subject to constant changes in land use (clearing, overgrazing, deforestation, and forest fires) that upset the hydrological balance. Through scenarios simulated with the Water Evaluation and Planning (WEAP) hydrological model, the present study analyzes the effects of forest fires and land use changes on the hydrological balance in the microwatersheds of central Mexico. The land use changes that took place between 1995 and 2021 were estimated, and projections based on the current scenario were made. Two trend scenarios were proposed for 2047: one with a positive trend (forest permanence) and the other with a negative trend (loss of cover from forest fires). The results show that with permanence or an increase in forest area, the surface runoff would decrease by 48.2%, increasing the base flow by 37% and the soil moisture by 2.3%. If forest is lost, surface runoff would increase up to 454%, and soil moisture would decrease by 27%. If the current forest decline trends continue, then there will be negative alterations in hydrological processes: a reduction in the interception of precipitation by the canopy and an increase in the velocity and flow of surface runoff, among others. The final result will be a lower amount of water being infiltrated into the soil and stored in the subsoil. The provision of hydrological environmental services depends on the maintenance of forest cover

Capacitive sensing of test mass motion with nanometer precision over millimeter-wide sensing gaps for space-borne gravitational reference sensors

We report on the performance of the capacitive gap-sensing system of the Gravitational Reference Sensor on board the LISA Pathfinder spacecraft. From in-flight measurements, the system has demonstrated a performance, down to 1 mHz, that is ranging between 0.7 and 1.8¿¿aF¿Hz-1/2. That translates into a sensing noise of the test mass motion within 1.2 and 2.4¿¿nm¿Hz-1/2 in displacement and within 83 and 170¿¿nrad¿Hz-1/2 in rotation. This matches the performance goals for LISA Pathfinder, and it allows the successful implementation of the gravitational waves observatory LISA. A 1/f tail has been observed for frequencies below 1 mHz, the tail has been investigated in detail with dedicated in-flight measurements, and a model is presented in the paper. A projection of such noise to frequencies below 0.1 mHz shows that an improvement of performance at those frequencies is desirable for the next generation of gravitational reference sensors for space-borne gravitational waves observation.Peer ReviewedPostprint (author's final draft

Biodegradable DFADs: Current status and prospects

Until recently, dFAD structure, materials and designs have remained quite rudimentary and virtually the same since their discovery, characterized by the increase of the dimensions and prevailing heavy use of plastic components. Biodegradable materials are called to be an important part of the solution, as they can faster degrade in the environment, free of toxins and heavy metals, reducing their lifespan, and preventing them from accumulating in sensitive areas once they are abandoned, lost or discarded. During last decades, regulatory measures at tRFMOs have advanced in the gradual implementation of biodegradable materials in dFAD constructions together with other measures limiting the number of active dFADs and the use of netting materials. However, more clarity is needed starting with a standardised definition of biodegradable dFADs among tRFMOs, to provide operational guidance. Research with those natural and synthetic materials is required, along with updated data collection for monitoring standards, as well as alternative and complementary actions need to be explored to contribute to minimising dFAD adverse effects on environment. Acknowledging the current difficulties for the implementation of fully biodegradable dFADs a stepwise process towards the implementation of fully biodegradable dFADs should be considered.Postprin