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Quantitative Analysis of Mitochondria-Associated Endoplasmic Reticulum Membrane (MAM) Stabilization in a Neural Model of Alzheimer's Disease (AD).
J Vis Exp
January 2025
Genetics and Aging Research Unit, MassGeneral Institute for Neurodegenerative Disease, Henry and Allison McCance Center for Brain Health, Department of Neurology, Massachusetts General Hospital, Harvard Medical School;
A method to quantitate the stabilization of Mitochondria-Associated endoplasmic reticulum Membranes (MAMs) in a 3-dimensional (3D) neural model of Alzheimer's disease (AD) is presented here. To begin, fresh human neuro progenitor ReN cells expressing β-amyloid precursor protein (APP) containing familial Alzheimer's disease (FAD) or naïve ReN cells are grown in thin (1:100) Matrigel-coated tissue culture plates. After the cells reach confluency, these are electroporated with expression plasmids encoding red fluorescence protein (RFP)-conjugated mitochondria-binding sequence of AKAP1(34-63) (Mito-RFP) that detects mitochondria or constitutive MAM stabilizers MAM 1X or MAM 9X that stabilize tight (6 nm ± 1 nm gap width) or loose (24 nm ± 3 nm gap width) MAMs, respectively.
View Article and Find Full Text PDFA novel ADP-directed chaperone function facilitates the ATP-driven motor activity of SARS-CoV helicase.
Nucleic Acids Res
January 2025
Single-Molecule and Cell Mechanobiology Laboratory, Daejeon, 34141, South Korea.
Helicase is a nucleic acid motor that catalyses the unwinding of double-stranded (ds) RNA and DNA via ATP hydrolysis. Helicases can act either as a nucleic acid motor that unwinds its ds substrates or as a chaperone that alters the stability of its substrates, but the two activities have not yet been reported to act simultaneously. Here, we used single-molecule techniques to unravel the synergistic coordination of helicase and chaperone activities, and found that the severe acute respiratory syndrome coronavirus helicase (nsp13) is capable of two modes of action: (i) binding of nsp13 in tandem with the fork junction of the substrate mechanically unwinds the substrate by an ATP-driven synchronous power stroke; and (ii) free nsp13, which is not bound to the substrate but complexed with ADP in solution, destabilizes the substrate through collisions between transient binding and unbinding events with unprecedented melting capability.
View Article and Find Full Text PDFCompeting addition processes give distinct growth regimes in the assembly of 1D filaments.
Biophys J
January 2025
Department of Physics, Kansas State University, Manhattan, KS 66506, USA. Electronic address:
We present a model to describe the concentration-dependent growth of protein filaments. Our model contains two states, a low entropy/high affinity ordered state and a high entropy/low affinity disordered state. Consistent with experiments, our model shows a diffusion-limited linear growth regime at low concentration, followed by a concentration-independent plateau at intermediate concentrations, and rapid disordered precipitation at the highest concentrations.
View Article and Find Full Text PDFHighly Permselective Contorted Polyamide Desalination Membranes with Enhanced Free Volume Fabricated by mLbL Assembly.
ACS Appl Mater Interfaces
January 2025
Civil and Environmental Engineering Department, University of Houston, 4226 Martin Luther King Blvd, Houston, Texas 77204, United States.
The permeability-selectivity trade-off in polymeric desalination membranes limits the efficiency and increases the costs of reverse osmosis and nanofiltration systems. Ultrathin contorted polyamide films with enhanced free volume demonstrate an impressive 8-fold increase in water permeance while maintaining equivalent salt rejection compared to conventional polyamide membranes made with -phenylenediamine and trimesoyl chloride monomers. The solution-based molecular layer-by-layer (mLbL) deposition technique employed for membrane fabrication sequentially reacts a shape-persistent contorted diamine monomer with a trimesoyl chloride monomer, forming highly cross-linked, dense polyamide networks while avoiding the kinetic and mass transfer limitations of traditional interfacial polymerization.
View Article and Find Full Text PDFCracking the code of acrylamide and Nε-(carboxymethyl)lysine: Fish oil use and predictive strategies in potato chips during thermal processing.
Food Chem
January 2025
National Engineering Laboratory of Intelligent Food Technology and Equipment, Zhejiang Key Laboratory of Agri-food Resources and High-value Utilization, Department of Food Science and Nutrition, College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou 310058, Zhejiang, China. Electronic address:
Global high consumption of fried potatoes is driven by appealing taste and edible convenience. However, the occurrence of Maillard reaction hazardous products (MRHPs) and joint control recipes have scarcely been concerned. We aim to reveal and predict how fish oil treatment for potato slices reduces simultaneous formation of typical MRHPs in air-based thermal processed potato chips.
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