Assessment of Mitochondrial Protein Topology and Membrane Insertion

Schäfer, Kerstin; Engstler, Carina; Dischinger, Korbinian; Carrie, Chris

doi:10.1007/978-1-0716-1653-6_13

Cited by 2 publications

(2 citation statements)

References 20 publications

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“…S1a-e), supporting the hypothesis of membrane rearrangements, in response to SoSe compared to vehicle-treated controls. To further validate these data, we determined the effects of SoSe-mediated mitophagy on the OMM localization of ICT1 31,32 and NUBP 33 (both IMM proteins) with/without proteinase K (PK) exposure 34 . ICT1 is a component of the large subunit of mitochondrial ribosomes, and NUBPL is a component of complex I, localized in the IMM.…”

Section: Resultsmentioning

confidence: 99%

Alterations of Lipid-Mediated Mitophagy Result in Aging-Dependent Sensorimotor Defects

Natalia

Albayram

Kassir

et al. 2022

Preprint

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The metabolic consequences of mitophagy alterations due to age-related stress in healthy aging brains vs. neurodegeneration remain unknown. Here, we demonstrate that ceramide synthase 1 (CerS1) is transported to the outer mitochondrial membrane (OMM) by the p17/PERMIT transporter that recognizes mislocalized mitochondrial ribosomes (mitoribosomes) via 39-FLRN-42 residues, inducing ceramide-mediated mitophagy. P17/PERMIT-CerS1-mediated mitophagy attenuated the argininosuccinate/fumarate/malate axis and induced D-glucose and fructose accumulation in neurons in culture, and brain tissues (primarily in the cerebellum) of wild-type mice in vivo. These metabolic changes in response to sodium-selenite were nullified in the cerebellum of CerS1top/top, PARKIN-/- or p17/PERMIT-/- mice that have dysfunctional mitophagy. Whereas sodium selenite induced mitophagy in the cerebellum, and improved motor-neuron deficits in aged wild-type mice, exogenous fumarate or malate prevented mitophagy. Attenuation of ceramide-mediated mitophagy enhanced damaged mitochondria accumulation and age-dependent sensorimotor abnormalities in p17/PERMIT-/- mice. Reinstituting mitophagy using a ceramide analog drug with selenium conjugate, LCL768, restored mitophagy and reduced malate/fumarate metabolism, improving sensorimotor deficits in old p17/PERMIT-/- mice. Thus, these data describe metabolic consequences of alterations to P17/PERMIT/ceramide-mediated mitophagy associated with the loss of mitochondrial quality control in the brain and provide therapeutic options to overcome age-dependent sensorimotor deficits and related disorders like amyotrophic lateral sclerosis (ALS).

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Section: Resultsmentioning

confidence: 99%

Alterations of Lipid-Mediated Mitophagy Result in Aging-Dependent Sensorimotor Defects

Natalia

Albayram

Kassir

et al. 2022

Preprint

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“…The optimized method allows the characterization of the CSPs and the identification of several hundreds of extracellular regions belonging to various transmembrane proteins (TMPs, the most important subclass of the CSPs that have at least one transmembrane segment). Although their high-resolution structural determinations are constantly evolving by emerging new biotechnological and artificial intelligence tools such as Cryo-EM and natural language models utilized by AlphaFold2 [38], other bioinformatical [39] and experimental methods [10,40] still have a great role in topology characterization of TMPs (topology defines the number of transmembrane segments and the orientation of the connecting loop relative to the membrane). While most of the existing experimental approaches often produce few topology data for only one TMP, the work presented here can produce several data for dozens of proteins at the same time.…”

Section: Introductionmentioning

confidence: 99%

Comprehensive Discovery of the Accessible Primary Amino Group-Containing Segments from Cell Surface Proteins by Fine-Tuning a High-Throughput Biotinylation Method

Langó

Kuffa

Tóth

et al. 2022

IJMS

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Cell surface proteins, including transmembrane and other surface-anchored proteins, play a key role in several critical cellular processes and have a strong diagnostic value. The development of quick and robust experimental methods remains vital for the accurate and comprehensive characterization of the cell surface subproteome of individual cells. Here we present a high-throughput technique which relies on the biotinylation of the accessible primary amino groups in the extracellular segments of the proteins, using HL60 as a model cell line. Several steps of the method have been thoroughly optimized to capture labeled surface proteins selectively and in larger quantities. These include the following: improving the efficiency of the cell surface biotinylation; reducing the endogen protease activity; applying an optimal amount of affinity column and elution steps for labeled peptide enrichment; and examining the effect of various solid-phase extraction methods, different HPLC gradients, and various tandem mass spectrometry settings. Using the optimized workflow, we identified at least 1700 surface-associated individual labeled peptides (~6000–7000 redundant peptides) from the model cell surface in a single nanoHPLC-MS/MS run. The presented method can provide a comprehensive and specific list of the cell surface available protein segments that could be potential targets in various bioinformatics and molecular biology research.

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Assessment of Mitochondrial Protein Topology and Membrane Insertion

Cited by 2 publications

References 20 publications

Alterations of Lipid-Mediated Mitophagy Result in Aging-Dependent Sensorimotor Defects

Alterations of Lipid-Mediated Mitophagy Result in Aging-Dependent Sensorimotor Defects

Comprehensive Discovery of the Accessible Primary Amino Group-Containing Segments from Cell Surface Proteins by Fine-Tuning a High-Throughput Biotinylation Method

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