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Neurodiversity refers to variations in the human brain that affect information processing; it includes conditions, or “neurotypes,” such as autism spectrum disorder (ASD), attention deficit hyperactivity disorder (ADHD), dyslexia, dyscalculia, and dyspraxia, among others. Neurodiversity can be conceptualized as significant differences in the ways that individuals process information; such differences may concern written or verbal language, sensory information, body language, or social interactions. These differences have been historically viewed within the medical model of disability, for example, as deficits in ability through a diagnosed condition, often associated with a goal of curing or managing the condition. 

Nuclear reactions play a crucial role in determining the nucleosynthesis that occurs in astrophysical events. The rates of many reactions that significantly impact certain nucleosynthesis processes can not be currently measured via direct means. These reactions must be constrained in another manner, such as determining the level energies and other structure properties of the compound nuclei. In order to measure level energies of nuclei relevant to nuclear astrophysics, the Enge split-pole spectrograph has been installed and commissioned at the University of Notre Dame’s Nuclear Science Laboratory. The first scientific measurement has also been performed. Structure properties of⁵⁸Cu were measured via the reaction⁵⁸Ni(³He,t)⁵⁸Cu to provide the first experimental constraint of the⁵⁷Ni(p,γ)⁵⁸Cu reaction rate, which impacts the production of of⁴⁴Ti,⁵⁷Fe, and⁵⁹Ni in core-collapse supernovae. Preliminary analysis of this measurement confirms the level energies of states in⁵⁸Cu that could lead to significant resonances in the⁵⁷Ni(p,γ)⁵⁸Cu reaction rate, while suggesting the presence of additional states that have not been previously observed but could also lead to significant resonances. 

With the recent explosion in high-resolution protein structures, one of the next frontiers in biology is elucidating the mechanisms by which conformational rearrangements in proteins are regulated to meet the needs of cells under changing conditions. Rigorously measuring protein energetics and dynamics requires the development of new methods that can resolve structural heterogeneity and conformational distributions. We have previously developed steady-state transition metal ion fluorescence resonance energy transfer (tmFRET) approaches using a fluorescent noncanonical amino acid donor (Anap) and transition metal ion acceptor to probe conformational rearrangements in soluble and membrane proteins. Here, we show that the fluorescent noncanonical amino acid Acd has superior photophysical properties that extend its utility as a donor for tmFRET. Using maltose-binding protein (MBP) expressed in mammalian cells as a model system, we show that Acd is comparable to Anap in steady-state tmFRET experiments and that its long, single-exponential lifetime is better suited for probing conformational distributions using time-resolved FRET. These experiments reveal differences in heterogeneity in the apo and holo conformational states of MBP and produce accurate quantification of the distributions among apo and holo conformational states at subsaturating maltose concentrations. Our new approach using Acd for time-resolved tmFRET sets the stage for measuring the energetics of conformational rearrangements in soluble and membrane proteins in near-native conditions. 

Acridonylalanine (Acd) is a fluorescent amino acid that is highly photostable, with a high quantum yield and long fluorescence lifetime in water. These properties make it superior to existing genetically encodable fluorescent amino acids for monitoring protein interactions and conformational changes through fluorescence polarization or lifetime experiments, including fluorescence lifetime imaging microscopy (FLIM). Here, we report the genetic incorporation of Acd using engineered pyrrolysine tRNA synthetase (RS) mutants that allow for efficient Acd incorporation in both E. coli and mammalian cells. We compare protein yields and amino acid specificity for these Acd RSs to identify an optimal construct. We also demonstrate the use of Acd in FLIM, where its long lifetime provides strong contrast compared to endogenous fluorophores and engineered fluorescent proteins, which have lifetimes less than 5 ns.