A5069434243- Sphingolipid Metabolism and Signaling
- Cellular transport and secretion
- Lipid Membrane Structure and Behavior
- Endoplasmic Reticulum Stress and Disease
- Carbohydrate Chemistry and Synthesis
- Fungal and yeast genetics research
- Polysaccharides and Plant Cell Walls
- Glycosylation and Glycoproteins Research
- Peptidase Inhibition and Analysis
- Plant-Microbe Interactions and Immunity
- Lipid metabolism and biosynthesis
- Lysosomal Storage Disorders Research
- Erythrocyte Function and Pathophysiology
- Photosynthetic Processes and Mechanisms
- Mitochondrial Function and Pathology
- ATP Synthase and ATPases Research
- Groundwater and Isotope Geochemistry
- Legume Nitrogen Fixing Symbiosis
- Environmental Justice and Health Disparities
- Biomedical Research and Pathophysiology
- Fungal Biology and Applications
- Hormonal Regulation and Hypertension
- Protein Tyrosine Phosphatases
- Landfill Environmental Impact Studies
- Phagocytosis and Immune Regulation
Kyushu University
2015-2025
Gifu University
2024-2025
Mukogawa Women's University
2021
Japan Science and Technology Agency
2010
Cambridge University Press
2010
New York University Press
2010
Hokkaido University
2003-2007
Medical University of South Carolina
2006-2007
Vanderbilt University
1996
University of Arizona
1992-1994
Previously, we reported two types of neutral ceramidase in mice, one solubilized by freeze-thawing and not. The former was purified as a 94-kDa protein from mouse liver, cloned (Tani, M., Okino, N., Mori, K., Tanigawa, T., Izu, H., Ito, M. (2000) <i>J. Biol. Chem.</i> 275, 11229–11234). In this paper, describe the purification, molecular cloning, subcellular distribution 112-kDa membrane-bound rat kidney, which completely insoluble freeze-thawing. open reading frame enzyme encoded...
The bioactive molecule sphingosine 1-phosphate (S1P) is abundantly stored in platelets and can be released extracellularly. However, although they have high (Sph) kinase activity, lack the de novo sphingolipid biosynthesis necessary to provide substrates. Here, we reveal a generation pathway for Sph, precursor of S1P, human platelets. Platelets incorporated extracellular 3H-labeled Sph much faster than megakaryoblastic cells rapidly converted it S1P. Furthermore, formed from plasma...
We report here a novel type of ceramidase of<i>Pseudomonas aeruginosa</i> AN17 isolated from the skin patient with atopic dermatitis. The enzyme was purified 83,400-fold an overall yield 21.1% culture supernatant strain AN17. After being stained silver staining solution, showed single protein band, and its molecular mass estimated to be 70 kDa on SDS-polyacrylamide gel electrophoresis. quite wide specificity for various ceramides,<i>i.e.</i> it hydrolyzed ceramides containing C12:0–C18:0...
We report here the molecular cloning, sequencing, and expression of gene encoding mouse neutral ceramidase, which has been proposed to function in sphingolipid signaling. A full-length cDNA ceramidase was cloned from a library liver using partial amino acid sequences purified ceramidase. The open reading frame 2,268 nucleotides encoded polypeptide 756 acids having nine putative N-glycosylation sites. Northern blot analysis revealed that mRNA expressed widely tissues, with especially strong...
Using C6-NBD-glucosylceramide (GlcCer) as a substrate, we detected the activity of conduritol B epoxide-insensitive neutral glycosylceramidase in cytosolic fractions zebrafish embryos, mouse and rat brains, human fibroblasts. The candidates for enzyme were assigned to Klotho (KL), whose family members share beta-glucosidase-like domain but natural substrates are unknown. Among this family, only KL-related protein (KLrP) is capable degrading C6-NBD-GlcCer when expressed CHOP cells, which...
We report here a novel ceramidase that was purified more than 150, 000-fold from the membrane fraction of mouse liver. The enzyme monomeric polypeptide having molecular mass 94 kDa and highly glycosylated with N-glycans. amino acid sequence fragment obtained homologous to those deduced genes encoding an alkaline Pseudomonas aeruginosa hypotheical protein slime mold Dictyostelium discoideum. However, no significant similarities were found in other known functional proteins including...
The neutral sphingomyelinases (nSMases) are considered major candidates for mediating the stress-induced production of ceramide. nSMase2, which has two hydrophobic segments near NH2-terminal region, been reported to be located at plasma membrane and play important roles in ceramide-mediated signaling. In this study, we found that nSMase2 is palmitoylated on multiple cysteine residues via thioester bonds. Site-directed mutagenesis alanine indicated clusters enzyme multiply palmitoylated; one...
Neutral sphingomyelinase 2 (nSMase2), which has two hydrophobic segments at its NH ‐terminus, plays an important role in ceramide‐mediated cell regulation. Here, we investigated the membrane topology of nSMase2. When a double‐tagged nSMase2 both and COOH termini, was overexpressed MCF‐7 cells, signals from tags were detected inner leaflet plasma membrane. Furthermore, insertion tag into internal sequence green fluorescent protein‐fused deletion mutants revealed that entire catalytic region...
In Saccharomyces cerevisiae, three classes of sphingolipids contain myo-inositol – inositol phosphorylceramide (IPC), mannosylinositol (MIPC) and mannosyldiinositol [M(IP)2C]. No fission yeast equivalent Ipt1p, the inositolphosphotransferase that synthesizes M(IP)2C from MIPC, has been found in Schizosaccharomyces pombe genome. Analysis sphingolipid composition wild-type cells confirmed MIPC is terminal most abundant complex S. pombe. Three proteins (Sur1p, Csg2p Csh1p) have shown to be...
Mitochondria of the yeast Saccharomyces cerevisiae contain enzymes Crd1p and Psd1p, which synthesize cardiolipin (CL) phosphatidylethanolamine respectively. A previous study indicated that crd1Δ is synthetically lethal with psd1Δ. In this study, to identify novel genes involved in CL metabolism, we searched for genetically interact found deletion FMP30 encoding a mitochondrial inner membrane protein results synthetic growth defect Although fmp30Δ cells grew normally exhibited slightly...
In the yeast Saccharomyces cerevisiae, complex sphingolipids have three types of polar head group, and breakdown their normal composition causes several cellular dysfunctions. Previously we found that loss biosynthesis mannosylinositol phosphorylceramide (MIPC) a defect in cell wall integrity (CWI). this study, screened for multicopy suppressor genes rescue CWI cells lacking MIPC synthases (Sur1 Csh1), is partly suppressed by upregulation ergosterol biosynthesis. addition, repression...
The budding yeast Saccharomyces cerevisiae can synthesise 15 subtypes of complex sphingolipids, and this structural diversity is thought to be the molecular basis that enables range biological functions sphingolipids. Through analyses mutants with various deletion combinations complex‐sphingolipid‐metabolising enzyme genes ( CSG1 , CSH1 IPT1 SUR2 SCS7 ), it was previously shown less sphingolipids leads increased sensitivity multiple environmental stresses, impaired plasma‐membrane cell‐wall...
Abstract Complex sphingolipids and sterols are coordinately involved in various cellular functions, e.g. the formation of lipid microdomains. Here we found that budding yeast exhibits resistance to an antifungal drug, aureobasidin A (AbA), inhibitor Aur1 catalyzing synthesis inositolphosphorylceramide, under impaired biosynthesis ergosterol, which includes deletion ERG6 , ERG2, or ERG5 final stages ergosterol pathway miconazole; however, these defects did not confer against repression...
Cryptococcus neoformans ( Cn ) is a fungal pathogen responsible for cryptococcal meningitis, which accounts 15% of AIDS-related deaths. Recent studies have shown that the absence sterol β-glucosidase (EGCrP2, also known as Sgl1) in significantly attenuates its virulence mouse infection model. However, mechanisms underlying this attenuation remain unclear. In study, we observed significant increase dead cells after 3 days culture SGL1 -deficient sgl1 Δ, KO) at 37°C, compared with wild-type...
We report here the molecular cloning and characterization of Drosophila neutral ceramidase (CDase). Using BLAST program, a CDase homologue (AE003774) was found in GenBank cloned from cDNA library imaginal discs. The open reading frame 2, 112 nucleo tides encoded polypeptide 704 amino acids having five putative N-glycosylation sites signal sequence composed 23 residues. When His-tagged overexpressed D. melanogaster Schneider's line 2 (S2) cells, enzyme continuously secreted into medium...
Neutral ceramidase is a type II integral membrane protein, which occasionally secreted into the extracellular milieu after processing of its N-terminal anchor. We found that when overexpressed in CHOP cells, neutral hydrolyzed cell surface ceramide, increased amount treatment cells with bacterial sphingomyelinase, leading to an increase cellular level sphingosine and 1-phosphate. On other hand, knockdown endogenous enzyme by siRNA decreased both sphingolipid metabolites. The bovine serum...