The acyl composition of sphingolipids is determined by the specificity of the enzyme ceramide synthase (EC 2.3.1.24). Ceramide contains a long-chain base (LCB) linked to a variety of fatty acids to produce a lipid class with potentially hundreds of structural variants. An optimized procedure for the assay of ceramide synthase in yeast microsomes is reported that uses mass spectrometry to detect any possible LCB and fatty acid combination synthesized from unlabeled substrates provided in the reaction. The assay requires the delivery of substrates with bovine serum albumin for maximum activity within defined limits of substrate concentration and specific methods to stop the reaction and extract the lipid that avoid the non-enzymatic synthesis of ceramide. The activity of ceramide synthase in yeast microsomes is demonstrated with the four natural LCBs found in yeast along with six saturated and two unsaturated fatty acyl-coenzyme As from 16 to 26 carbons in length. The procedure allows for the determination of substrate specificity and kinetic parameters toward natural substrates for ceramide synthase from potentially any organism.
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http://dx.doi.org/10.1016/j.ab.2015.02.016 | DOI Listing |
Physiol Plant
January 2025
Laboratory of Plant Physiology, Universidad de Extremadura, Badajoz, Spain.
Plant sphingolipids are lipophilic membrane components essential for different cellular functions but they also act as signaling molecules in various aspects of plant development. However, the interaction between plant sphingolipids and abscission remains largely uncharacterized. Here, the possible role of sphingolipids in regulating fruit abscission was examined in the abscission zone (AZ) of olive fruit.
View Article and Find Full Text PDFToxicology
January 2025
Department of Occupational Health and Environmental Health, School of Public Health, Anhui Medical University, Hefei, Anhui 230032, PR China. Electronic address:
Bisphenol A (BPA) is a typical environmental endocrine disruptor which have been broadly confirmed to be associated with malignant tumors, including colorectal cancer (CRC). Lipid metabolism reprogramming performed important biological effects in cancer progression. While the role of lipid metabolism in CRC progression upon BPA exposure remain elusive.
View Article and Find Full Text PDFMech Ageing Dev
December 2024
School of Medicine, Nankai University, 94 Weijin Road, Tianjin 30071, China; Institude of Chinese PLA Geriatric Medicine, The Second Medical Centre, Chinese PLA General Hospital & National Clinical Research Center for Geriatric Diseases, Beijing 100853, China. Electronic address:
Objective: The senescence of smooth muscle is one of the independent risk factors in atherosclerosis progression in which the vascular inflammation plays an important role on vascular dysfunction. This study is designed to explore the novel vascular aging biomarkers and screen the potential molecular interventional targets through bioinformatic analysis.
Results: Transcriptional analysis was conducted based on the GSE16487 open access database, which included 15 human vascular tissue samples from two groups: young group (≤ 60 years old, n = 8) and aged group (≥ 75 years old, n = 7).
mBio
December 2024
(Epi-)Genetic Regulation of Fungal Virulence, Leibniz Institute for Natural Product Research and Infection Biology-Hans Knöll Institute (Leibniz-HKI), Jena, Germany.
produces the mycotoxin fumonisin B (FB), which disrupts sphingolipid biosynthesis by inhibiting ceramide synthase and affects the health of plants, animals, and humans. The means by which protects itself from its own mycotoxin are not completely understood. Some fumonisin () cluster genes do not contribute to the biosynthesis of the compound, but their function has remained enigmatic.
View Article and Find Full Text PDFFoods
November 2024
Department of Medical Nutrition, Kyung Hee University, Yongin 17104, Republic of Korea.
This study investigated the protective effects of a ceramides derivates from the peach (PF3) on photoaging by UV-irradiated hairless mice. Mice were randomly divided into seven groups: AIN93G without UVB exposure (normal control, NC), AIN93G with UVB exposure (control, C), AIN93G supplemented 100 mg/kg body weight (BW) of L-ascorbic acid with UVB exposure (AA), AIN93G supplemented 100 mg/kg BW of arbutin with UVB exposure (Arbutin), AIN93G supplemented 10 mg/kg BW of PF3 with UVB exposure (10PF3), AIN93G supplemented 20 mg/kg BW of PF3 with UVB exposure (20PF3), and AIN93G supplemented 40 mg/kg BW of PF3 with UVB exposure (40PF3). The study examined the impact of PF3 on skin hydration, wrinkle formation, and melanogenesis using enzyme-linked immunosorbent assay (ELISA), real-time polymerase chain reaction (real-time PCR), and Western blot analysis.
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