Methods for detection of lateral domains by solid-state 2H nuclear magnetic resonance (NMR) and 1H magic angle spinning (MAS)-NMR in model- and biomembranes are presented. 2H NMR has been used for decades to distinguish between liquid-ordered and solid-ordered lamellar phases of phospholipids with deuterated hydrocarbon chains. More recently, it was shown that superposition of liquid-ordered and -disordered phases is detected as well, taking advantage of the large differences in chain order parameters between them. Experiments require preparation of samples with deuterated lipids. In contrast, 1H MAS-NMR utilizes the natural proton NMR signals of lipids in model- and biomembranes. Very good resolution of resonances according to their chemical shifts is achieved by rapid spinning of samples at the "magic angle" (54.7 degrees) to the main magnetic field. Phase transitions to ordered states are detected as broadening of resonances. The method distinguishes liquid-disordered, liquid-ordered, and solid-ordered phases, has much higher sensitivity than 2H NMR, and does not require labeling. In combination with pulsed magnetic field gradients, 1H MAS-NMR yields diffusion rates that may report confinement of lipids to domains with submicrometer dimensions.
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http://dx.doi.org/10.1007/978-1-59745-513-8_9 | DOI Listing |
Hum Genet
January 2025
Department of Biomedical Sciences, University of Padova, Padova, Italy.
The Genetics of Neurodevelopmental Disorders Lab in Padua provided a new intellectual disability (ID) Panel challenge for computational methods to predict patient phenotypes and their causal variants in the context of the Critical Assessment of the Genome Interpretation, 6th edition (CAGI6). Eight research teams submitted a total of 30 models to predict phenotypes based on the sequences of 74 genes (VCF format) in 415 pediatric patients affected by Neurodevelopmental Disorders (NDDs). NDDs are clinically and genetically heterogeneous conditions, with onset in infant age.
View Article and Find Full Text PDFInt J Mol Sci
December 2024
Department of Biosciences, Biotechnology and Environment, University of Bari Aldo Moro, Via E. Orabona 4, 70125 Bari, Italy.
Mitochondrial carriers transport organic acids, amino acids, nucleotides and cofactors across the mitochondrial inner membrane. These transporters consist of a three-fold symmetric bundle of six transmembrane α-helices that encircle a pore with a central substrate binding site, whose alternating access is controlled by a cytoplasmic and a matrix gate (C- and M-gates). The C- and M-gates close by forming two different salt-bridge networks involving the conserved motifs [YF][DE]XX[KR] on the even-numbered and PX[DE]XX[KR] on the odd-numbered transmembrane α-helices, respectively.
View Article and Find Full Text PDFInt J Mol Sci
December 2024
Department of Biosciences, Biotechnology and Environment, University of Bari Aldo Moro, 70125 Bari, Italy.
Mutations in the SLC25A38 gene are responsible for the second most common form of congenital sideroblastic anemia (CSA), a severe condition for which no effective treatment exists. We developed and characterized a K562 erythroleukemia cell line with markedly reduced expression of the SLC25A38 protein (A38-low cells). This model successfully recapitulated the main features of CSA, including reduced heme content and mitochondrial respiration, increase in mitochondrial iron, ROS levels and sensitivity to oxidative stress.
View Article and Find Full Text PDFLife (Basel)
December 2024
Division of Microbiology and Molecular Biology, Medical Faculty, Private Sigmund Freud University, Freudplatz 3, 1020 Wien, Austria.
The attractiveness of OMVs derived from Gram-negative bacteria lies in the fact that they have two biomembranes sandwiching a peptidoglycan layer. It is well known that the envelope of OMVs consists of the outer bacterial membrane [OM] and not of the inner one [IM] of the source bacterium. This implies that all outer membranous molecules found in the OM act as antigens.
View Article and Find Full Text PDFProc Natl Acad Sci U S A
January 2025
Institute of Optical Materials and Chemical Biology, Guangxi Key Laboratory of Electrochemical Energy Materials, School of Chemistry and Chemical Engineering, Guangxi University, Nanning 530004, Guangxi, People's Republic of China.
Monitoring subcellular organelle dynamics in real time and precisely assessing membrane heterogeneity in living cells are very important for studying fundamental biological mechanisms and gaining a comprehensive understanding of cellular processes. However, there remains a shortage of effective tools for these purposes. Herein, we propose a strategy to develop the exchangeable water-sensing probeAPBD for time-lapse imaging of dynamics in cellular membrane-bound organelle morphology with structured illumination microscopy at the nanoscale.
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