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https://www.readbyqxmd.com/read/29092960/quantification-of-3-ketodihydrosphingosine-using-hplc-esi-ms-ms-to-study-spt-activity-in-yeast-saccharomyces-cerevisiae
#1
Jihui Ren, Justin Snider, Michael V Airola, Aaron Zhong, Nadia A Rana, Lina M Obeid, Yusuf A Hannun
Serine palmitoyltransferase (SPT) catalyzes the rate-limiting step of condensation of L-serine and palmitoyl-CoA to form 3-ketodihydrosphingosine (3KDS). Here we report a HPLC-ESI-MS/MS method to directly quantify 3KDS generated by SPT. With this technique we were able to detect 3KDS at a level comparable to that of dihydrosphingosine (DHS) in yeast S.cerevisiae. An in vitro SPT assay measuring the incorporation of deuterated serine into deuterated 3KDS was developed. The results show that SPT kinetics in response to palmitoyl-CoA fit into an allosteric sigmoidal model, suggesting the existence of more than one palmitoyl-CoA binding site on yeast SPT and positive cooperativity between them...
November 1, 2017: Journal of Lipid Research
https://www.readbyqxmd.com/read/29025713/neutrophil-elastase-increases-airway-ceramide-levels-via-upregulation-of-serine-palmitoyltransferase
#2
Sophia Karandashova, Apparao B Kummarapurugu, Shuo Zheng, Charles Chalfant, Judith A Voynow
Altered sphingolipid metabolism is associated with increased inflammation; however, the impact of inflammatory mediators, including neutrophil elastase (NE), on airway sphingolipid homeostasis remains unknown. Using a well-characterized mouse model of NE oropharyngeal aspiration, we investigated a potential link between NE-induced airway inflammation and increased synthesis of various classes of sphingolipids, including ceramide species. Sphingolipids in bronchoalveolar lavage fluids (BAL) were identified and quantified using reverse phase high-performance liquid chromatography/electrospray ionization tandem mass spectrometry analysis...
October 12, 2017: American Journal of Physiology. Lung Cellular and Molecular Physiology
https://www.readbyqxmd.com/read/28857357/aspergillus-nidulans-baph-is-a-rab11-effector-that-connects-membranes-in-the-spitzenk%C3%A3-rper-with-basal-autophagy
#3
Mario Pinar, Miguel A Peñalva
Using affinity chromatography we identified the Aspergillus nidulans F-BAR-and-PH domain-containing protein BapH as a RabE(RAB11) effector. BapH localizes to the Spitzenkörper (SPK) in an F-actin- and Sec7-dependent manner, becoming cytosolic after inactivation of Trs120 in TRAPPII, the oligomeric GEF for RabE(RAB11) . Therefore, RabE(RAB11) contributes to the recruitment of BapH to secretory vesicles in vivo. BapH has a close homologue, SlmA, which is related to yeast Slm1p/Slm2p, localizes to eisosomes and does not bind RabE(RAB11) ...
November 2017: Molecular Microbiology
https://www.readbyqxmd.com/read/28714922/application-of-an-improved-hplc-fl-method-to-screen-serine-palmitoyl-transferase-inhibitors
#4
Simone Bertini, Giuseppe Saccomanni, Sara Del Carlo, Maria Digiacomo, Claudia Gargini, Ilaria Piano, Giuseppe Matteo Campisi, Riccardo Ghidoni, Marco Macchia, Clementina Manera
In this work, we reported the application and validation of an improved high-performance liquid chromatography method coupled with a fluorimetric detector (HPLC-FL) to screen the activity of two heterocyclic derivatives reported as serine palmitoyl transferase (SPT) inhibitors. The analytical conditions were optimized in terms of the derivatization procedure, chromatographic condition, extraction procedure, and method validation according to EMEA guidelines. Once fully optimized, the method was applied to assess the SPT-inhibitory activity of the above-mentioned derivatives and of the reference inhibitor myriocin...
July 17, 2017: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
https://www.readbyqxmd.com/read/28699686/ablation-of-ceramide-synthase-2-exacerbates-dextran-sodium-sulphate-induced-colitis-in-mice-due-to-increased-intestinal-permeability
#5
Ye-Ryung Kim, Giora Volpert, Kyong-Oh Shin, So-Yeon Kim, Sun-Hye Shin, Younghay Lee, Sun Hee Sung, Yong-Moon Lee, Jung-Hyuck Ahn, Yael Pewzner-Jung, Woo-Jae Park, Anthony H Futerman, Joo-Won Park
Ceramides mediate crucial cellular processes including cell death and inflammation and have recently been implicated in inflammatory bowel disease. Ceramides consist of a sphingoid long-chain base to which fatty acids of various length can be attached. We now investigate the effect of alerting the ceramide acyl chain length on a mouse model of colitis. Ceramide synthase (CerS) 2 null mice, which lack very-long acyl chain ceramides with concomitant increase of long chain bases and C16-ceramides, were more susceptible to dextran sodium sulphate-induced colitis, and their survival rate was markedly decreased compared with that of wild-type littermates...
July 12, 2017: Journal of Cellular and Molecular Medicine
https://www.readbyqxmd.com/read/28679588/adipocyte-mtorc1-deficiency-promotes-adipose-tissue-inflammation-and-nlrp3-inflammasome-activation-via-oxidative-stress-and-de-novo-ceramide-synthesis
#6
Patricia Chimin, Maynara L Andrade, Thiago Belchior, Vivian A Paschoal, Juliana Magdalon, Alex S Yamashita, Érique Castro, Angela Castoldi, Adriano B Chaves-Filho, Marcos Y Yoshinaga, Sayuri Miyamoto, Niels O Câmara, William T Festuccia
Mechanistic target of rapamycin complex (mTORC)1 activity is increased in adipose tissue of obese insulin-resistant mice, but its role in the regulation of tissue inflammation is unknown. Herein, we investigated the effects of adipocyte mTORC1 deficiency on adipose tissue inflammation and glucose homeostasis. For this, mice with adipocyte raptor deletion and controls fed a chow or a high-fat diet were evaluated for body mass, adiposity, glucose homeostasis, and adipose tissue inflammation. Despite reducing adiposity, adipocyte mTORC1 deficiency promoted hepatic steatosis, insulin resistance, and adipose tissue inflammation (increased infiltration of macrophages, neutrophils, and B lymphocytes; crown-like structure density; TNF-α, interleukin (IL)-6, and monocyte chemoattractant protein 1 expression; IL-1β protein content; lipid peroxidation; and de novo ceramide synthesis)...
September 2017: Journal of Lipid Research
https://www.readbyqxmd.com/read/28674344/total-synthesis-of-sphingofungin-e-and-4-5-di-epi-sphingofungin-e
#7
Narumi Noda, Hisanori Nambu, Tomoya Fujiwara, Takayuki Yakura
Total synthesis of sphingofungin E and 4,5-di-epi-sphingofungin E was achieved from an intermediate same as that of myriocin and mycestericin D via antipodal stereoselective dihydroxylations.
2017: Chemical & Pharmaceutical Bulletin
https://www.readbyqxmd.com/read/28621883/determination-of-the-serine-palmitoyl-transferase-inhibitor-myriocin-by-electrospray-and-q-trap-mass-spectrometry
#8
Giuseppe Matteo Campisi, Paola Signorelli, Jessica Rizzo, Claudio Ghilardi, Jacopo Antognetti, Anna Caretti, Jelena S Lazarević, Enrica Strettoi, Elena Novelli, Riccardo Ghidoni, Federico Maria Rubino, Rita Paroni
Myriocin is a potent inhibitor of serine-palmitoyl-transferase, the first and rate-determining enzyme in the sphingolipids biosynthetic pathway. This study developed, validated and applied a LC-MS/MS method to measure myriocin in minute specimens of animal tissue. The chemical analog 14-OH-myriocin was used as the internal standard. The two molecules were extracted from the tissue homogenate by solid-phase extraction, separated by gradient reversed-phase liquid chromatography and measured by negative ion electrospray mass spectrometry in the triple quadrupole...
June 16, 2017: Biomedical Chromatography: BMC
https://www.readbyqxmd.com/read/28600071/synthetic-antimicrobial-peptides-of-the-halictines-family-disturb-the-membrane-integrity-of-candida-cells
#9
Marie Kodedová, Hana Sychrová
We compared the potency of four derivatives of the antimicrobial peptide halictine-2 against six Candida species. Observed activity was peptide and species specific. Halictines rapidly permeabilized cell membranes and caused the leakage of cytosolic components. Their killing potential was enhanced by the commercial antimicrobial agent octenidine dihydrochloride. The effect on C. glabrata cells did not depend on the activity of Cdr pumps, but was influenced by their lipid composition. The pre-treatment of cells with myriocin, an inhibitor of sphingolipid synthesis, enhanced the peptides' activity, whereas pre-treatment with terbinafine and fluconazole, inhibitors of sterol synthesis, significantly weakened their efficacy...
October 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28531105/high-mobility-group-box-1-disrupts-metabolic-function-with-cigarette-smoke-exposure-in-a-ceramide-dependent-manner
#10
Oliver J Taylor, Mikayla O Thatcher, Sheryl T Carr, Jonathan L Gibbs, Annie M Trumbull, Mitchell E Harrison, Duane R Winden, Mackenzie J Pearson, Trevor S Tippetts, William L Holland, Paul R Reynolds, Benjamin T Bikman
We have previously found that cigarette smoke disrupts metabolic function, in part, by increasing muscle ceramide accrual. To further our understanding of this, we sought to determine the role of the cytokine high-mobility group box 1 (HMGB1), which is increased with smoke exposure, in smoke-induced muscle metabolic perturbations. To test this theory, we determined HMGB1 from lungs of human smokers, as well as from lung cells from mice exposed to cigarette smoke. We also treated cells and mice directly with HMGB1, in the presence or absence of myriocin, an inhibitor of serine palmitoyltransferase, the rate-limiting enzyme in ceramide biosynthesis...
May 20, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28490444/inhibiting-glucosylceramide-synthase-exacerbates-cisplatin-induced-acute-kidney-injury
#11
Tess V Dupre, Mark A Doll, Parag P Shah, Cierra N Sharp, Deanna Siow, Judit Megyesi, James Shayman, Alicja Bielawska, Jacek Bielawski, Levi J Beverly, Maria Hernandez-Corbacho, Christopher J Clarke, Ashley J Snider, Rick G Schnellmann, Lina M Obeid, Yusuf A Hannun, Leah J Siskind
Acute kidney injury (AKI), resulting from chemotherapeutic agents such as cisplatin, remains an obstacle in the treatment of cancer. Cisplatin-induced AKI involves apoptotic and necrotic cell death, pathways regulated by sphingolipids such as ceramide and glucosylceramide. Results from this study indicate that C57BL/6J mice treated with cisplatin had increased ceramide and hexosylceramide levels in the renal cortex 72 h following cisplatin treatment. Pretreatment of mice with inhibitors of acid sphingomyelinase and de novo ceramide synthesis (amitriptyline and myriocin, respectively) prevented accumulation of ceramides and hexosylceramide in the renal cortex and protected from cisplatin-induced AKI...
July 2017: Journal of Lipid Research
https://www.readbyqxmd.com/read/28474276/sphingosine-toxicity-in-eae-and-ms-evidence-for-ceramide-generation-via-serine-palmitoyltransferase-activation
#12
Lawrence G Miller, Jennifer A Young, Swapan K Ray, Guanghu Wang, Sharad Purohit, Naren L Banik, Somsankar Dasgupta
Multiple sclerosis (MS) is a demyelinating disorder characterized by massive neurodegeneration and profound axonal loss. Since myelin is enriched with sphingolipids and some of them display toxicity, biological function of sphingolipids in demyelination has been investigated in MS brain tissues. An elevation of sphingosine with a decrease in monoglycosylceramide and psychosine (myelin markers) was observed in MS white matter and plaque compared to normal brain tissue. This indicated that sphingosine toxicity might mediate oligodendrocyte degeneration...
May 5, 2017: Neurochemical Research
https://www.readbyqxmd.com/read/28469091/increased-de-novo-ceramide-synthesis-and-accumulation-in-failing-myocardium
#13
Ruiping Ji, Hirokazu Akashi, Konstantinos Drosatos, Xianghai Liao, Hongfeng Jiang, Peter J Kennel, Danielle L Brunjes, Estibaliz Castillero, Xiaokan Zhang, Lily Y Deng, Shunichi Homma, Isaac J George, Hiroo Takayama, Yoshifumi Naka, Ira J Goldberg, P Christian Schulze
Abnormal lipid metabolism may contribute to myocardial injury and remodeling. To determine whether accumulation of very long-chain ceramides occurs in human failing myocardium, we analyzed myocardial tissue and serum from patients with severe heart failure (HF) undergoing placement of left ventricular assist devices and controls. Lipidomic analysis revealed increased total and very long-chain ceramides in myocardium and serum of patients with advanced HF. After unloading, these changes showed partial reversibility...
May 4, 2017: JCI Insight
https://www.readbyqxmd.com/read/28439630/myriocin-treatment-of-cf-lung-infection-and-inflammation-complex-analyses-for-enigmatic-lipids
#14
Anna Caretti, Michele Vasso, Fabiola Tecla Bonezzi, Andrea Gallina, Marco Trinchera, Alice Rossi, Raffaella Adami, Josefina Casas, Monica Falleni, Delfina Tosi, Alessandra Bragonzi, Riccardo Ghidoni, Cecilia Gelfi, Paola Signorelli
Our aim was to use quantitative and qualitative analyses to gain further insight into the role of ceramide in cystic fibrosis (CF). Sphingolipid ceramide is a known inflammatory mediator, and its accumulation in inflamed lung has been reported in different types of emphysema, chronic obstructive pulmonary disease and CF. CF is caused by a mutation of the chloride channel and associated with hyperinflammation of the respiratory airways and high susceptibility to ongoing infections. We have previously demonstrated that de novo ceramide synthesis is enhanced in lung inflammation and sustains Pseudomonas aeruginosa pulmonary infection in a CF murine model...
August 2017: Naunyn-Schmiedeberg's Archives of Pharmacology
https://www.readbyqxmd.com/read/28409208/inhibitors-of-ceramide-de-novo-biosynthesis-rescue-damages-induced-by-cigarette-smoke-in-airways-epithelia
#15
Aida Zulueta, Anna Caretti, Giuseppe Matteo Campisi, Andrea Brizzolari, Jose Luis Abad, Rita Paroni, Paola Signorelli, Riccardo Ghidoni
Exposure to cigarette smoke represents the most important risk factor for the development of chronic obstructive pulmonary disease (COPD). COPD is characterized by chronic inflammation of the airways, imbalance of proteolytic activity resulting in the destruction of lung parenchyma, alveolar hypoxia, oxidative stress, and apoptosis. Sphingolipids are structural membrane components whose metabolism is altered during stress. Known as apoptosis and inflammation inducer, the sphingolipid ceramide was found to accumulate in COPD airways and its plasma concentration increased as well...
July 2017: Naunyn-Schmiedeberg's Archives of Pharmacology
https://www.readbyqxmd.com/read/28268212/inhibition-of-ceramide-decreased-the-expression-of-atp-binding-cassette-transporter-g5-8-mrna-in-an-animal-model-of-cholesterol-gallstone
#16
Hyo Jung Kim, Jae Seon Kim, Seikwan Oh, Hwan-Soo Yoo
BACKGROUND: The increased risk of gallstone has been reported in patients with ATP-binding cassette (ABC) transporter polymorphism. The half-transporters ABCG5 and ABCG8 mediate the efflux of cholesterol in hepatocytes and the intestine. We investigated whether ceramide plays a role in cholesterol efflux through the ABC transporters. METHODS: Six-week-old C57BL/6J mice were assigned to 3 groups. The normal group (n = 5) was fed a normal chow diet, the cholesterol group (n = 10) was fed a lithogenic diet, and the myriocin group (n = 15) was fed the lithogenic diet and myriocin, a specific inhibitor of serine-palmitoyl transferase...
2017: Digestive Diseases
https://www.readbyqxmd.com/read/28236661/fam20c-is-under-the-control-of-sphingolipid-signaling-in-human-cell-lines
#17
Giorgio Cozza, Mauro Salvi, Vincent S Tagliabracci, Lorenzo A Pinna
Fam20C, also termed DMP-4 (dentin matrix protein 4) and G-CK (Golgi casein kinase) is an atypical protein kinase committed with the phosphorylation of casein and a plethora of other secreted proteins. Fam20C has been implicated in a number of human pathologies related to biomineralization, phosphate homeostasis, and neoplasia. The mode of regulation of Fam20C is still a matter of conjecture. In in vitro, Fam20C activity is stimulated several fold by sphingosine. To gain in vivo information about the physiological relevance of this observation, three cell lines expressing endogenous Fam20C, and one in which Fam20C has been knocked out with CRISPR/Cas9 technology have been examined for Fam20C activity under basal conditions and where sphingosine has been depleted by treatment with myriocin...
April 2017: FEBS Journal
https://www.readbyqxmd.com/read/28189121/myriocin-treatment-affects-lipid-metabolism-in-skeletal-muscles-of-rats-with-streptozotocin-induced-type-1-diabetes
#18
Krzysztof Kurek, Marta Garbowska, Dominika M Ziembicka, Bartłomiej Łukaszuk, Jakub Rogowski, Adrian Chabowski, Jan Górski, Małgorzata Żendzian-Piotrowska
PURPOSE: The aim of this work was to assess the effect(s) of de novo ceramide synthesis inhibition on lipid metabolism in skeletal muscle tissue of type 1 diabetic rats. The latter seems to be of vital importance, since previous works have shown its positive influence on lipid metabolism and glucose homeostasis in the case of its counterpart - type 2 diabetes. MATERIALS/METHODS: The animals were randomly assigned to one of the following groups: C - control, M - myriocin (ceramide de novo synthesis inhibitor), D - diabetes (induced by streptozotocin injections); D+M - diabetes+myriocin...
March 2017: Advances in Medical Sciences
https://www.readbyqxmd.com/read/28152593/combining-deep-sequencing-proteomics-phosphoproteomics-and-functional-screens-to-discover-novel-regulators-of-sphingolipid-homeostasis
#19
Nicolas Lebesgue, Márton Megyeri, Alba Cristobal, Arjen Scholten, Silvia G Chuartzman, Yoav Voichek, Richard A Scheltema, Shabaz Mohammed, Anthony H Futerman, Maya Schuldiner, Albert J R Heck, Simone Lemeer
Sphingolipids (SLs) are essential components of cell membranes and are broad-range bioactive signaling molecules. SL levels must be tightly regulated as imbalances affect cellular function and contribute to pathologies ranging from neurodegenerative and metabolic disorders to cancer and aging. Deciphering how SL homeostasis is maintained and uncovering new regulators is required for understanding lipid biology and for identifying new targets for therapeutic interventions. Here we combine omics technologies to identify the changes of the transcriptome, proteome, and phosphoproteome in the yeast Saccharomyces cerevisiae upon SL depletion induced by myriocin...
February 3, 2017: Journal of Proteome Research
https://www.readbyqxmd.com/read/28123341/downregulation-of-lipin-1-induces-insulin-resistance-by-increasing-intracellular-ceramide-accumulation-in-c2c12-myotubes
#20
Shujuan Huang, Suling Huang, Xi Wang, Qingli Zhang, Jia Liu, Ying Leng
Dysregulation of lipid metabolism in skeletal muscle is involved in the development of insulin resistance. Mutations in lipin-1, a key lipid metabolism regulator leads to significant systemic insulin resistance in fld mice. However, the function of lipin-1 on lipid metabolism and insulin sensitivity in skeletal muscle is still unclear. Herein we demonstrated that downregulation of lipin-1 in C2C12 myotubes by siRNA transfection suppressed insulin action, characterized by reduced insulin stimulated Akt phosphorylation and glucose uptake...
2017: International Journal of Biological Sciences
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