Results for “RAS” — Page 4

DOE OSTI2021

Intercomparison of MJO Column Moist Static Energy and Water Vapor Budget among Six Modern Reanalysis Products

This study conducts an intercomparison of the column-integrated moist static energy (MSE) and water vapor budget of the Madden–Julian oscillation (MJO) among six modern global reanalysis products (RAs). Inter-RA differences in the mean MSE, MJO MSE anomalies, individual MSE budget terms, and their relative contributions to the propagation and maintenance of MJO MSE anomalies are examined. Also investigated is the relationship between the MJO column water vapor (CWV) budget residuals with the other CWV budget terms as well as with the two parameters that characterize cloud–radiation feedback and moisture–convection coupling. Results show a noticeable inter-RA spread in the mean-state MSE, especially its vertical structure. In all RAs, horizontal MSE advection dominates the propagation of the MJO MSE while column-integrated longwave radiative heating and vertical MSE advection are found to be the key processes for MJO maintenance. The MSE budget terms directly affected by the model parameterization schemes exhibit high uncertainty. The differences in anomalous vertical velocity mainly contribute to the large differences in vertical MSE advection among the RAs. The budget residuals show large inter-RA differences and have nonnegligible contributions to MJO maintenance and propagation in most RAs. RAs that underestimate (overestimate) the strength of cloud–radiation feedback and the convective moisture adjustment time scale tend to have positive (negative) MJO CWV budget residual, indicating the critical role of these processes in the maintenance of MJO CWV anomalies. Our results emphasize that a correct representation of the interactions among moisture, convection, cloud, and radiation is the key for an accurate depiction of the MJO MSE and CWV budget in RAs.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2021

Deep Learning-Based Adaptive Remedial Action Scheme with Security Margin for Renewable-Dominated Power Grids

The Remedial Action Scheme (RAS) is designed to take corrective actions after detecting predetermined conditions to maintain system transient stability in large interconnected power grids. However, since RAS is usually designed based on a few selected typical operating conditions, it is not optimal in operating conditions that are not considered in the offline design, especially under frequently and dramatically varying operating conditions due to the increasing integration of intermittent renewables. The deep learning-based RAS is proposed to enhance the adaptivity of RAS to varying operating conditions. During the training, a customized loss function is developed to penalize the negative loss and suggest corrective actions with a security margin to avoid triggering under-frequency and over-frequency relays. Simulation results of the reduced United States Western Interconnection system model demonstrate that the proposed deep learning–based RAS can provide optimal corrective actions for unseen operating conditions while maintaining a sufficient security margin.

24 POWER TRANSMISSION AND DISTRIBUTION↗

NASA NTRS1999

Hypertrophic response to hemodynamic overload: role of load vs. renin-angiotensin system activation

Myocardial hypertrophy is one of the basic mechanisms by which the heart compensates for hemodynamic overload. The mechanisms by which hemodynamic overload is transduced by the cardiac muscle cell and translated into cardiac hypertrophy are not completely understood. Candidates include activation of the renin-angiotensin system (RAS) and angiotensin II receptor (AT1) stimulation. In this study, we tested the hypothesis that load, independent of the RAS, is sufficient to stimulate cardiac growth. Four groups of cats were studied: 14 normal controls, 20 pulmonary artery-banded (PAB) cats, 7 PAB cats in whom the AT1 was concomitantly and continuously blocked with losartan, and 8 PAB cats in whom the angiotensin-converting enzyme (ACE) was concomitantly and continuously blocked with captopril. Losartan cats had at least a one-log order increase in the ED50 of the blood pressure response to angiotensin II infusion. Right ventricular (RV) hypertrophy was assessed using the RV mass-to-body weight ratio and ventricular cardiocyte size. RV hemodynamic overload was assessed by measuring RV systolic and diastolic pressures. Neither the extent of RV pressure overload nor RV hypertrophy that resulted from PAB was affected by AT1 blockade with losartan or ACE inhibition with captopril. RV systolic pressure was increased from 21 +/- 3 mmHg in normals to 68 +/- 4 mmHg in PAB, 65 +/- 5 mmHg in PAB plus losartan and 62 +/- 3 mmHg in PAB plus captopril. RV-to-body weight ratio increased from 0.52 +/- 0.04 g/kg in normals to 1.11 +/- 0.06 g/kg in PAB, 1.06 +/- 0.06 g/kg in PAB plus losartan and 1.06 +/- 0.06 g/kg in PAB plus captopril. Thus 1) pharmacological modulation of the RAS with losartan and captopril did not change the extent of the hemodynamic overload or the hypertrophic response induced by PAB; 2) neither RAS activation nor angiotensin II receptor stimulation is an obligatory and necessary component of the signaling pathway that acts as an intermediary coupling load to the hypertrophic response; and 3) load, independent of the RAS, is capable of stimulating cardiac growth.

Non-NASA Center↗

NASA NTRSDate not supplied

Advanced Air Mobility Operations & Automation Part II Technical Lecture

The Airborne Collision Avoidance System X (ACAS X) is a next-generation collision avoidance tool developed to support different aircraft types and operations. The rotorcraft variant – referred to as ACAS XR – is designed to accommodate existing helicopter platforms as well as in-development, electric vertical takeoff and landing concepts, which are critical to the emerging concept of operations referred to as Advanced Air Mobility. The fundamental role of ACAS XR is to provide Detect and Avoid (DAA) and/or Collision Avoidance (CA) protection against airborne traffic. DAA alerting and guidance in the context of ACAS XR is caution-level and “suggestive,” and is to be used by the pilot if, and when, they decide to maneuver against an identified threat to DAA “well clear.” The CA alerting, by contrast, is warning-level and “directive,” with the pilot required to comply with the associated guidance to prevent a predicted Near Midair Collision (NMAC). The CA alerts generated by ACAS XR are referred to as Resolution Advisories (RAs) consistent with previous CA systems. Unlike earlier CA systems, ACAS XR issues RAs in the horizontal and vertical dimensions as well as multi-axis RAs (referred to as “Blended” RAs). According to the Minimal Operational Performance Standards of DAA systems for Unmanned Aircraft Systems, maneuvers to comply with RAs may be automated, whereas maneuvers based on DAA alerting assume a manual response. The current presentation discusses two human-in-the-loop simulations that presented rotorcraft pilots with ACAS XR alerts and guidance in eVTOL simulators with varying levels of automation. It also presents overviews of these studies as well as how they assisted live flight tests, which will occur throughout 2023.

air taxis↗

DOE OSTI2021

Transformative remedial action scheme tool (TRAST)

Techniques and apparatuses are described that enable transformative Remedial Action Scheme (RAS) analyses and methodologies for a bulk electric power system, including methods of designing, reviewing, revising, testing, implementing, verifying, or validating a RAS. An improved RAS improves operation of the power system, including performance, reliability, control, and asset utilization. The example methodologies discussed—also referred to as a transformative Remedial Action Scheme tool (TRAST)—provide an end-to-end solution for adaptively setting RAS parameters based on realistic and near real-time operation conditions to improve power grid reliability and grid asset utilization, by leveraging utility data analysis and employing dynamic simulations and machine learning to significantly simplify and shorten the entire RAS process.

Fan, Xiaoyuan↗

DOE OSTI2024

The CNK–HYP scaffolding complex promotes RAF activation by enhancing KSR–MEK interaction

The RAS–MAPK pathway regulates cell proliferation, differentiation and survival, and its dysregulation is associated with cancer development. The pathway minimally comprises the small GTPase RAS and the kinases RAF, MEK and ERK. Activation of RAF by RAS is notoriously intricate and remains only partially understood. There are three RAF isoforms in mammals (ARAF, BRAF and CRAF) and two related pseudokinases (KSR1 and KSR2). RAS-mediated activation of RAF depends on an allosteric mechanism driven by the dimerization of its kinase domain. Recent work on human RAFs showed that MEK binding to KSR1 promotes KSR1–BRAF heterodimerization, which leads to the phosphorylation of free MEK molecules by BRAF. Similar findings were made with the single Drosophila RAF homolog. Here we show that the fly scaffold proteins CNK and HYP stabilize the KSR–MEK interaction, which in turn enhances RAF–KSR heterodimerization and RAF activation. The cryogenic electron microscopy structure of the minimal KSR–MEK–CNK–HYP complex reveals a ring-like arrangement of the CNK–HYP complex allowing CNK to simultaneously engage KSR and MEK, thus stabilizing the binary interaction. Together, these results illuminate how CNK contributes to RAF activation by stimulating the allosteric function of KSR and highlight the diversity of mechanisms impacting RAF dimerization as well as the regulatory potential of the KSR–MEK interaction.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2024

Membrane lipids drive formation of KRAS4b-RAF1 RBDCRD nanoclusters on the membrane

The oncogene RAS, extensively studied for decades, presents persistent gaps in understanding, hindering the development of effective therapeutic strategies due to a lack of precise details on how RAS initiates MAPK signaling with RAF effector proteins at the plasma membrane. Recent advances in X-ray crystallography, cryo-EM, and super-resolution fluorescence microscopy offer structural and spatial insights, yet the molecular mechanisms involving protein-protein and protein-lipid interactions in RAS-mediated signaling require further characterization. This study utilizes single-molecule experimental techniques, nuclear magnetic resonance spectroscopy, and the computational Machine-Learned Modeling Infrastructure (MuMMI) to examine KRAS4b and RAF1 on a biologically relevant lipid bilayer. MuMMI captures long-timescale events while preserving detailed atomic descriptions, providing testable models for experimental validation. Both in vitro and computational studies reveal that RBDCRD binding alters KRAS lateral diffusion on the lipid bilayer, increasing cluster size and decreasing diffusion. RAS and membrane binding cause hydrophobic residues in the CRD region to penetrate the bilayer, stabilizing complexes through β-strand elongation. These cooperative interactions among lipids, KRAS4b, and RAF1 are proposed as essential for forming nanoclusters, potentially a critical step in MAP kinase signal activation.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2020

Mechanistic insights explain the transforming potential of the T507K substitution in the protein-tyrosine phosphatase SHP2

The protein-tyrosine phosphatase SHP2 is an allosteric enzyme critical for cellular events downstream of growth factor receptors. Mutations in the SHP2 gene have been linked to many different types of human diseases, including developmental disorders, leukemia, and solid tumors. Unlike most SHP2-activating mutations, the T507K substitution in SHP2 is unique in that it exhibits oncogenic Ras-like transforming activity. However, the biochemical basis of how the SHP2/T507K variant elicits transformation remains unclear. By combining kinetic and biophysical methods, X-ray crystallography, and molecular modeling, as well as using cell biology approaches, in this work we uncovered that the T507K substitution alters both SHP2 substrate specificity and its allosteric regulatory mechanism. We found that although SHP2/T507K exists in the closed, autoinhibited conformation similar to the WT enzyme, the interactions between its N-SH2 and protein-tyrosine phosphatase domains are weakened such that SHP2/T507K possesses a higher affinity for the scaffolding protein Grb2-associated binding protein 1 (Gab1). We also discovered that the T507K substitution alters the structure of the SHP2 active site, resulting in a change in SHP2 substrate preference for Sprouty1, a known negative regulator of Ras signaling and a potential tumor suppressor. Our results suggest that SHP2/T507K's shift in substrate specificity coupled with its preferential association of SHP2/T507K with Gab1 enable the mutant SHP2 to more efficiently dephosphorylate Sprouty1 at pTyr-53. This dephosphorylation hyperactivates Ras signaling, which is likely responsible for SHP2/T507K's Ras-like transforming activity.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2020

Investigation of Automated Corrective Actions for Special Protection Schemes

The constantly evolving nature of the grid is compelling the design process of Remedial Action Schemes (RAS) to keep up with the changes. This document proposes a flexible and computationally efficient approach to automatically determine RAS corrective actions that alleviate line overloading violations. Statistical and functional characteristics summarized from RAS implemented in real power systems are used to guide the design parameters. This report also leverages sensitivity-based techniques to determine corrective actions for specific contingencies quickly without repeated numerical simulations. Finally, future directions for implementing this approach for a fully automated, online RAS are discussed.

24 POWER TRANSMISSION AND DISTRIBUTION↗

NASA NTRS2016

Impact of Flight Enthalpy, Fuel Simulant, and Chemical Reactions on the Mixing Characteristics of Several Injectors at Hypervelocity Flow Conditions

The high total temperatures or total enthalpies required to duplicate the high-speed flight conditions in ground experiments often place stringent requirements on the material selection and cooling needs for the test articles and intrusive flow diagnostic equipment. Furthermore, for internal flows, these conditions often complicate the use of nonintrusive diagnostics that need optical access to the test section and interior portions of the flowpath. Because of the technical challenges and increased costs associated with experimentation at high values of total enthalpy, an attempt is often made to reduce it. This is the case for the Enhanced Injection and Mixing Project (EIMP) currently underway in the Arc-Heated Scramjet Test Facility at the NASA Langley Research Center. The EIMP aims to investigate supersonic combustion ramjet (scramjet) fuel injection and mixing physics, improve the understanding of underlying physical processes, and develop enhancement strategies and functional relationships between mixing performance and losses relevant to flight Mach numbers greater than 8. The experiments will consider a "direct-connect" approach and utilize a Mach 6 nozzle to simulate the combustor entrance flow of a scramjet engine. However, while the value of the Mach number is matched to that expected at the combustor entrance in flight, the maximum value of the total enthalpy for these experiments is limited by the thermal-structural limits of the uncooled experimental hardware. Furthermore, the fuel simulant is helium, not hydrogen. The use of "cold" flows and non-reacting mixtures of fuel simulants for mixing experiments is not new and has been extensively utilized as a screening technique for scramjet fuel injectors. In this study, Reynolds-averaged simulations are utilized (RAS) to systematically verify the implicit assumptions used by the EIMP. This is accomplished by first performing RAS of mixing for two injector configurations at planned nominal experimental conditions. The mixing parameters of interest, such as mixing efficiency and total pressure recovery, are then computed and compared to the values obtained from RAS under the true enthalpy conditions and using helium and hydrogen. Finally, the impact of combustion on mixing, often deemed small enough to neglect at hypervelocity conditions, is assessed by comparing the results obtained from the hydrogen-fueled reacting and non-reacting RAS. For reacting flows, in addition to mixing efficiency and total pressure recovery, the combustion efficiency and thrust potential are also considered. In all of the simulations, the incoming air Mach number and the fuel-to-air ratio are the same, while the total pressure, total enthalpy, and the fuel simulant vary depending on the case considered. It is found that under some conditions the "cold" flow experiments are a good approximation of the flight.

Drozda, Tomasz G.↗

NASA NTRS2019

Numerical Investigation and Optimization of a Flushwall Injector for Scramjet Applications at Hypervelocity Flow Conditions

An investigation utilizing Reynolds-averaged simulations (RAS) was performed in order to demonstrate the use of design and analysis of computer experiments (DACE) methods in Sandia’s DAKOTA software package for surrogate modeling and optimization. These methods were applied to a flow- path fueled with an interdigitated flushwall injector suitable for scramjet applications at hyper- velocity conditions and ascending along a constant dynamic pressure flight trajectory. The flight Mach number, duct height, spanwise width, and injection angle were the design variables selected to maximize two objective functions: the thrust potential and combustion efficiency. Because the RAS of this case are computationally expensive, surrogate models are used for optimization. To build a surrogate model a RAS database is created. The sequence of the design variables comprising the database were generated using a Latin hypercube sampling (LHS) method. A methodology was also developed to automatically build geometries and generate structured grids for each design point. The ensuing RAS analysis generated the simulation database from which the two objective functions were computed using a one-dimensionalization (1D) of the three-dimensional simulation data. The data were fitted using four surrogate models: an artificial neural network (ANN), a cubic polynomial, a quadratic polynomial, and a Kriging model. Variance-based decomposition showed that both objective functions were primarily driven by changes in the duct height. Multiobjective design optimization was performed for all four surrogate models via a genetic algorithm method. Optimal solutions were obtained at the upper and lower bounds of the flight Mach number range. The Kriging model predicted an optimal solution set that exhibited high values for both objective functions. Additionally, three challenge points were selected to assess the designs on the Pareto fronts. Further sampling among the designs of the Pareto fronts may be required to lower the surrogate model errors and perform more accurate surrogate-model-based optimization.

Shenoy, Rajiv R.↗

NASA NTRSDate not supplied

A Human-in-the-Loop Evaluation of ACAS Xu

As part of the Phase 2 UAS DAA MOPS, a Class 3 DAA system has been under development with the potential to resolve many of the limitations of Class 1 and 2 systems. A Class 3 system would combine the DAA and CA functions into a single, unified system, and would also extend the CA capabilities relative to TCAS II. Class 3 is enabled by the Airborne Collision Avoidance System (ACAS) XU, a next generation CA system developed specifically for UAS operations. Unlike Class 2 systems, ACAS XU provides CA protection against both cooperative and non-cooperative traffic. Class 3 systems also expand the CA logic to allow horizontal RAs in addition to vertical RAs. The purpose of the current study was to evaluate ACAS XU in a real-time, HITL simulation. The latest version of ACAS XU was implemented and pilots were tasked with responding to scripted traffic conflicts over the course of four experimental trials. Variables included the location of the ACAS XU guidance information (standalone vs. integrated) and traffic conflict type (DAA or CA threat). Sixteen active UAS pilots participated in the study, with ATC and ‘pseudo’ pilots acting as airspace confederates. Results showed that pilots were able to maintain DWC with ACAS XU at a rate comparable to previous research (~5%). Compliance rates to initial RAs were high (~90%) but dropped significantly when the target heading value issued during horizontal RAs were updated over the course of an encounter (30-70%). The implications of these findings on the display, alerting, and guidance requirements for Class 3 systems will be discussed.

unmanned aircraft systems↗

NASA NTRSDate not supplied

A Human-in-the-Loop Evaluation of ACAS Xu

As part of the Phase 2 UAS DAA MOPS, a Class 3 DAA system has been under development with the potential to resolve many of the limitations of Class 1 and 2 systems. A Class 3 system would combine the DAA and CA functions into a single, unified system, and would also extend the CA capabilities relative to TCAS II. Class 3 is enabled by the Airborne Collision Avoidance System (ACAS) XU, a next generation CA system developed specifically for UAS operations. Unlike Class 2 systems, ACAS XU provides CA protection against both cooperative and non-cooperative traffic. Class 3 systems also expand the CA logic to allow horizontal RAs in addition to vertical RAs. The purpose of the current study was to evaluate ACAS XU in a real-time, HITL simulation. The latest version of ACAS XU was implemented and pilots were tasked with responding to scripted traffic conflicts over the course of four experimental trials. Variables included the location of the ACAS XU guidance information (standalone vs. integrated) and traffic conflict type (DAA or CA threat). Sixteen active UAS pilots participated in the study, with ATC and ‘pseudo’ pilots acting as airspace confederates. Results showed that pilots were able to maintain DWC with ACAS XU at a rate comparable to previous research (~5%). Compliance rates to initial RAs were high (~90%) but dropped significantly when the target heading value issued during horizontal RAs were updated over the course of an encounter (30-70%). The implications of these findings on the display, alerting, and guidance requirements for Class 3 systems will be discussed.

unmanned aircraft systems↗

NASA NTRS2020

A Human-in-theLoop Evaluation of ACAS Xu

As part of the Phase 2 UAS DAA MOPS, a Class 3 DAA system has been under development with the potential to resolve many of the limitations of Class 1 and 2 systems. A Class 3 system would combine the DAA and CA functions into a single, unified system, and would also extend the CA capabilities relative to TCAS II. Class 3 is enabled by the Airborne Collision Avoidance System (ACAS) XU, a next generation CA system developed specifically for UAS operations. Unlike Class 2 systems, ACAS XU provides CA protection against both cooperative and non-cooperative traffic. Class 3 systems also expand the CA logic to allow horizontal RAs in addition to vertical RAs. The purpose of the current study was to evaluate ACAS XU in a real-time, HITL simulation. The latest version of ACAS XU was implemented and pilots were tasked with responding to scripted traffic conflicts over the course of four experimental trials. Variables included the location of the ACAS XU guidance information (standalone vs. integrated) and traffic conflict type (DAA or CA threat). Sixteen active UAS pilots participated in the study, with ATC and ‘pseudo’ pilots acting as airspace confederates. Results showed that pilots were able to maintain DWC with ACAS XU at a rate comparable to previous research (~5%). Compliance rates to initial RAs were high (~90%) but dropped significantly when the target heading value issued during horizontal RAs were updated over the course of an encounter (30-70%). The implications of these findings on the display, alerting, and guidance requirements for Class 3 systems will be discussed.

unmanned aircraft systems↗

NASA NTRSDate not supplied

Understanding the Role of CFD, Including RANS, DES, LES and DNS, in Hypersonic Propulsion Design and Analysis

Computational fluid dynamics (CFD) has matured to the point where it is becoming ubiquitous in the design and analysis of aerospace vehicles and their propulsion systems. The current state-of-the-art of CFD in hypersonic propulsion design and analysis as well as a historical perspective of its development over the last 50 years is presented. The current state-of-the-art for the design of finite rate chemically reacting turbulent hypersonic flow is shown to currently be Reynolds averaged simulations (RAS) where the Reynolds averaged Navier-Stokes (RAS) equations are solved to obtain steady state solutions. An overview of the physical modeling techniques employed by RAS for the simulation of hypersonic chemically reacting flow are presented. However, scale resolving simulation techniques, such as detached eddy simulation (DES), large eddy simulations (LES), hybrid RAS/LES and direct numerical simulations (DNS), are becoming becoming powerful tools for point design analysis. One example of such a scale resolving technique, hybrid RAS/LES, will be briefly described as well as how it can be used to help address model form uncertainties in RAS.

Hypersonic↗

DOE OSTI2022

Lifetime of actin-dependent protein nanoclusters

Protein nanoclusters (PNCs) are dynamic collections of a few proteins that spatially organize in nanometer-length clusters. PNCs are one of the principal forms of spatial organization of membrane proteins, and they have been shown or hypothesized to be important in various cellular processes, including cell signaling. PNCs show remarkable diversity in size, shape, and lifetime. In particular, the lifetime of PNCs can vary over a wide range of timescales. The diversity in size and shape can be explained by the interaction of the clustering proteins with the actin cytoskeleton or the lipid membrane, but very little is known about the processes that determine the lifetime of the nanoclusters. In this paper, using mathematical modeling of the cluster dynamics, we model the biophysical processes that determine the lifetime of actin-dependent PNCs. In particular, we investigated the role of actin aster fragmentation, which had been suggested to be a key determinant of the PNC lifetime, and we found that it is important only for a small class of PNCs. A simple extension of our model allowed us to investigate the kinetics of protein-ligand interaction near PNCs. We found an anomalous increase in the lifetime of ligands near PNCs, which agrees remarkably well with experimental data on RAS-RAF kinetics. In particular, analysis of the RAS-RAF data through our model provides falsifiable predictions and novel hypotheses that will not only shed light on the role of RAS-RAF kinetics in various cancers, but also will be useful in studying membrane protein clustering in general.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2020

Mobility of Radionuclides in Fractured Carbonate Rocks: Lessons from a Field-Scale Transport Experiment

Current research on radionuclide disposal is mostly conducted in granite, clay, saltstone, or volcanic tuff formations. These rock types are not always available to host a geological repository in every nuclear waste-generating country, but carbonate rocks may serve as a potential alternative. To assess their feasibility, a forced gradient cross-borehole tracer experiment was conducted in a saturated fractured chalk formation. The mobility of stable Sr and Cs (as analogs for their radioactive counterparts), Ce (an actinide analog), Re (a Tc analog), bentonite particles, and fluorescent dye tracers through the flow path was analyzed. The migration of each of these radionuclide analogs (RAs) was shown to be dependent upon their chemical speciation in solution, their interactions with bentonite, and their sorption potential to the chalk rock matrix. The brackish groundwater resulted in flocculation and immobilization of most particulate RAs. Nevertheless, the high permeability of the fracture system allowed for fast overall transport times of all aqueous RAs investigated. This study suggests that the geochemical properties of carbonate rocks may provide suitable conditions for certain types of radionuclide storage (in particular, brackish, high-porosity, and low-permeability chalks). Nevertheless, careful consideration should be given to high-permeability fracture networks that may result in high radionuclide mobility.

38 RADIATION CHEMISTRY, RADIOCHEMISTRY, AND NUCLEA↗

DOE OSTI2026

Free Energy and Flexibility Analysis of Autoinhibited Human BRAF

The RAF serine/threonine protein kinases function as direct effectors of RAS in the intracellular transmission of extracellular growth signals, and they are key targets for drug discovery, given the high incidence of oncogenic mutations in RAF and other components of this signaling pathway. In its inactive state, RAF is held in an autoinhibited conformation in the cytosol through a combination of intramolecular interactions and binding to a regulatory 14−3−3 protein dimer. Activation of RAF is initiated by its interaction with membrane-localized GTP-bound RAS, which induces conformational changes that release RAF from its autoinhibited state. However, the molecular mechanisms governing RAF activation remain incomplete, largely due to the challenges in experimentally capturing the intermediate conformational states in this process. To address this gap, we developed a comprehensive all-atom model of BRAF based on existing cryo-EM structures. Using this model, we performed extensive molecular dynamics simulations to evaluate the stability and free energy landscape of autoinhibited BRAF in solution. Our analysis reveals conformational flexibility within the autoinhibited complex, suggesting that this dynamic behavior may play a role in facilitating BRAF activation upon engagement with the membrane-bound RAS.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗