Two alternative specific and very sensitive methods for determination of vinyl epoxide synthetase activity in liver microsomes are described. Trichloroethylene, which is used as a substrate, is converted into trichloroethylene oxide by a hepatic epoxide synthetase. Chloral hydrate, the final rearrangement product of trichloroethylene oxide, is evaluated by electron capture gas chromatography, either after derivatization with pentafluorophenyl-hydrazine or after its conversion into chloroform under alcaline conditions. The kinetic parameters of the epoxidative reaction have been determined on rat hepatic microsomal suspensions.
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http://dx.doi.org/10.1007/978-3-642-66896-8_60 | DOI Listing |
Physiol Rev
April 2025
Institute of Metabolism and Cell Death, Molecular Targets and Therapeutics Center, Helmholtz Zentrum München, Neuherberg, Germany.
We present here a comprehensive update on recent advancements in the field of ferroptosis, with a particular emphasis on its metabolic underpinnings and physiological impacts. After briefly introducing landmark studies that have helped to shape the concept of ferroptosis as a distinct form of cell death, we critically evaluate the key metabolic determinants involved in its regulation. These include the metabolism of essential trace elements such as selenium and iron; amino acids such as cyst(e)ine, methionine, glutamine/glutamate, and tryptophan; and carbohydrates, covering glycolysis, the citric acid cycle, the electron transport chain, and the pentose phosphate pathway.
View Article and Find Full Text PDFMethods Enzymol
November 2024
Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC, United States. Electronic address:
Gamma-glutamyl carboxylase (GGCX), a polytopic membrane protein found in the endoplasmic reticulum, catalyzes the posttranslational modification of a variety of vitamin K-dependent (VKD) proteins to their functional forms. GGCX uses the free energy from the oxygenation of reduced vitamin K to remove the proton from the glutamate residue to drive VKD carboxylation. During the process of carboxylation, reduced vitamin K is oxidized to vitamin K epoxide.
View Article and Find Full Text PDFMethods Enzymol
November 2024
Department of Inflammation and Immunity, Lerner Research Institute, Cleveland Clinic Lerner College of Medicine at CWRU, Cleveland, OH, United States.
Angew Chem Int Ed Engl
December 2024
School of Chemistry, University of Bristol, BS8 1TS, Bristol, UK.
The delineation of the complex biosynthesis of the potent antibiotic mupirocin, which consists of a mixture of pseudomonic acids (PAs) isolated from Pseudomonas fluorescens NCIMB 10586, presents significant challenges, and the timing and mechanisms of several key transformations remain elusive. Particularly intriguing are the steps that process the linear backbone from the initial polyketide assembly phase to generate the first cyclic intermediate PA-B. These include epoxidation as well as incorporation of the tetrahydropyran (THP) ring and fatty acid side chain required for biological activity.
View Article and Find Full Text PDFJ Lipid Res
September 2024
Division of Bioorganic Chemistry and Molecular Pharmacology, Department of Medicine, Washington University School of Medicine, Saint Louis, MO, USA; Department of Chemistry, Washington University, Saint Louis, MO, USA; Department of Developmental Biology, Washington University School of Medicine, Saint Louis, MO, USA; Center for Cardiovascular Research, Washington University School of Medicine, Saint Louis, MO, USA. Electronic address:
Mitochondrial fatty acid oxidation serves as an essential process for cellular survival, differentiation, proliferation, and energy metabolism. Numerous studies have utilized etomoxir (ETO) for the irreversible inhibition of carnitine palmitoylcarnitine transferase 1 (CPT1), which catalyzes the rate-limiting step for mitochondrial long-chain fatty acid β-oxidation to examine the bioenergetic roles of mitochondrial fatty acid metabolism in many tissues in multiple diverse disease states. Herein, we demonstrate that intact mitochondria robustly metabolize ETO to etomoxir-carnitine (ETO-carnitine) prior to nearly complete ETO-mediated inhibition of CPT1.
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