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| 1 | Human intestinal acyl-CoA synthetase 5 is sensitive to the inhibitor triacsin C显示文摘AIM:To investigate whether human acyl-CoA synthetase 5(ACSL5) is sensitive to the ACSL inhibitor triacsin C.METHODS:The ACSL isoforms ACSL1 and ACSL5 from rat as well as human ACSL5 were cloned and recombinantly expressed as 6xHis-tagged enzymes.Ni 2+-affinity purified recombinant enzymes were assayed at pH 7.5 or pH 9.5 in the presence or absence of triacsin C.In addition,ACSL5 transfected CaCo2 cells and intestinal human mucosa were monitored.ACSL5 expression in cellular systems was verified using Western blot and immunofluorescence.The ACSL assay mix included TrisHCl(pH 7.4),ATP,CoA,EDTA,DTT,MgCl 2,[9,103 H] palmitic acid,and triton X-100.The 200 μL reaction was initiated with the addition of solubilized,purified recombinant proteins or cellular lysates.Reactions were terminated after 10,30 or 60 min of incubation with Doles medium.RESULTS:Expression of soluble recombinant ACSL proteins was found after incubation with isopropyl betaD-1-thiogalactopyranoside and after ultracentrifugation these were further purified to near homogeneity with Ni 2+-affinity chromatography.Triacsin C selectively and strongly inhibited recombinant human ACSL5 protein at pH 7.5 and pH 9.5,as well as recombinant rat ACSL1(sensitive control),but not recombinant rat ACSL5(insensitive control).The IC50 for human ACSL5 was about 10 μmol/L.The inhibitory triacsin C effect was similar for different incubation times(10,30 and 60 min) and was not modified by the N-or C-terminal location of the 6xHis-tag.In order to evaluate ACSL5 sensitivity to triacsin C in a cellular environment,stable human ACSL5 CaCo2 transfectants and mechanically dissected normal human intestinal mucosa with high physiological expression of ACSL5 were analyzed.In both models,ACSL5 peak activity was found at pH 7.5 and pH 9.5,corresponding to the properties of recombinant human ACSL5 protein.In the presence of triacsin C(25 μmol/L),total ACSL activity was dramatically diminished in human ACSL5 transfectants as well as in ACSL5-rich human intestinal mucosa.CONCLUSION:The data strongly indicate that human ACSL5 is sensitive to triacsin C and does not compensate for other triacsin C-sensitive ACSL isoforms. | Elke Kaemmerer Anne Peuscher Andrea Reinartz Christian Liedtke Ralf Weiskirchen Jürgen Kopitz Nikolaus Gassler | 2011 | World Journal of Gastroenterology2011，17，44： | 3 |
| 2 | Lipid-induced up-regulation of human acyl-CoA synthetase 5 promotes hepatocellular apoptosis显示文摘 | Andrea Reinartz Josef Ehling Andrea Leue Christian Liedtke Ursula Schneider Jürgen Kopitz Thomas Weiss Claus Hellerbrand Ralf Weiskirchen Ruth Knüchel Nikolaus Gassler | 2010 | BBA - Molecular and Cell Biology of Lipids2010，，9： | 2 |
| 3 | Lipid peroxidation products reduce lysosomal protease activities in human retinal pigment epithelial cells via two different mechanisms of action显示文摘 | Tim U. Krohne Elke Kaemmerer Frank G. Holz Jürgen Kopitz | 2009 | Experimental Eye Research2009，，2： | 1 |
| 4 | Negative regulation of neuroblastoma cell growth by carbohydrate-dependent surface binding of galectin-1 and functional divergence from galectin-3显示文摘 | Kopitz J von Reitzenstein C Gabius HJ | 2001 | J Biol Chem2001，276，35： | 1 |
| 5 | Photodamage to human RPE ceils by A2-E, a retinoid component of lipofuscin 显示文摘 | Schtitt F Davies S Kopitz J | 2000 | Invest OphthalmoI Vis Sei2000，41，： | 1 |
| 6 | Tyrosinase biosynthesis in adult mammalian retinal pigment epithelial cells显示文摘 | Schraermeyer U Kopitz J Peters S | 2006 | Experimental Eye Research2006，83，： | 1 |
| 7 | Plasma amisulpride levels in schizophrenia or schizoaffective disorder显示文摘 | Bergemann N Kopitz J Kress KP Frick A | 2004 | Eur Neuropsychopharmacol2004，14，3： | 1 |
| 8 | Insulin-resistant brain state after intracerebroventricular steptozocin injection exacerbates Alzhe- imer-like changes in Tg2576 Al~PP-overexpressing mice 显示文摘 | Plaschke K Kopitz J Sieqelin M | 2010 | J Alzheimers Dis2010，19，： | 1 |
| 9 | Effects of cell surface ganglioside sialidase inhibition on growth control and differentiation of human neuroblastoma cells 显示文摘 | Kopitz J Muhl C Ehemann V | 1997 | Eur J Cell Biol1997，73，1： | 1 |
| 10 | Proteins modified by malondialdehyde, 4-hydroxynonenal, or advanced glycation end products in lipofuscin of human retinal pigment epithelium显示文摘 | Schutt F Bergmann M Holz FG Kopitz J | 2003 | Invest Ophthalmol Vis Sci2003，44，8： | 1 |
| 11 | Homodimeric ga-lectin-7 (p53-induced gene 1) is a negative growth regulator for human neuroblastoma cells显示文摘 | KOPITZ J ANDRE S VON R C | 2003 | Oncogene2003，22，40： | 1 |
| 12 | Heat shock proteins as gatekeepers of proteolytic pathways-implications for age-related macular degeneration (AMD) 显示文摘 | Kaarniranta K Salminen A Eskelinen EL Kopitz J | 2009 | Ageing Res Rev2009，8，2： | 1 |
| 13 | Proteins modified by malondialdehyde,4-hydroxynonenal,or advanced glycation end products in lipofuscin of human retinal pigment epithelium显示文摘 | Schutt F Bergmann M Holz FG Kopitz J | 2003 | Invest Ophthalmol Vis Sci2003，44，8： | 1 |
| 14 | Olanzapine plasma concentration, average daily dose, and interaction with co-medication in schizophrenic patients显示文摘 | Bergemann N Frick A Parzer P Kopitz J | 2004 | Pharmacopsychiatry2004，37，2： | 1 |
| 15 | Lipids and lipid peroxidation products in the pathogenesis of age-related macular degeneration 显示文摘 | Kopitz J Holz FG Kaemmerer E Schutt F | 2004 | Biochemic2004，6，11： | 1 |
| 16 | Insulin-resistant brain stateafter intracerebroventricular streptozotocin injection exacerbates Al-zheimer-like changes in Tg2576 AbetaPP-overexpressing mice 显示文摘 | Plaschke K Kopitz J Siegelin M | 2010 | J Alzheimers Dis2010，19，2： | 1 |
| 17 | Insulin-resistant brain state after intracerebroventricular streptozotocin injection exacerbates Alzheimer-hke changes in Tg2576 AbetaPP- overexpressing mice显示文摘 | Plaschke K Kopitz J Siegelin M | 2010 | J Alzheimers Dis2010，19，2： | 1 |
| 18 | Proteome analysis of lipofuscin in human retinal pigment epithelial cells显示文摘 | Schutt F Ueberle B Schnolzer M Holz FG Kopitz J | 2002 | FEBS Lett2002，528，： | 1 |
| 19 | Proteins modified by malondialdehyde,4-hydroxynonenal,or advanced glycation end products in lipofuscin of human retinal pigment epithelium显示文摘 | Schutt F Bergmann M Holz FG Kopitz J | 2003 | Invest Ophthalmol Vis Sci2003，44，： | 1 |
| 20 | Inhibition of lysosomal degradative functions in RPE cells by a retinoid component of lipofuscin 显示文摘 | HOLZ FG SCHUTT F KOPITZ J ELDRWD GE KRUSE FE VOLCKER HE | 1999 | Invest Ophthalmol Vis Sci1999，40，3： | 1 |
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