Synthesis of a novel fluorescent ceramide analogue and its use in the characterization of recombinant ceramidase from Pseudomonas aeruginosa PA01

Chemistry and Physics of Lipids
W F NieuwenhuizenMaarten R Egmond

Abstract

Ceramidase (CDase) hydrolyses the N-acyl linkage of the sphingolipid ceramide. We synthesized the non-fluorescent ceramide analogue (4E,2S,3R)-2-N-(10-pyrenedecanoyl)-1,3,17-trihydroxy-17-(3,5-dinitrobenzoyl)-4-heptadecene (10) that becomes fluorescent upon hydrolysis of its N-acyl bond. This novel substrate was used to study several kinetic aspects of the recombinant CDase from the pathogenic bacterium Pseudomonas aeruginosa PA01. Maximum CDase activity was observed above 1.5 microM substrate, with an apparent K(m) of 0.5+/-0.1 microM and a turnover of 5.5 min(-1). CDase activity depends on divalent cations without a strong specificity. CDase is inhibited by sphingosine and by several sphingosine analogues. The lack of inhibition by several mammalian CDase inhibitors such as D-erythro-MAPP, L-erythro-MAPP or N-oleoylethanolamine points to a novel active site and/or substrate binding region. The CDase assay described here offers the opportunity to develop and screen for specific bacterial CDase inhibitors of pharmaceutical interest.

References

Jul 30, 1990·FEBS Letters·P W Wertz, D T Downing
May 1, 1986·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·M W SpenceH W Cook
May 12, 1995·The Journal of Biological Chemistry·K BernardoK Sandhoff
May 24, 1996·The Journal of Biological Chemistry·A BielawskaY A Hannun
Jun 11, 1998·The Journal of Biological Chemistry·N OkinoM Ito
Jan 1, 1999·Bioorganic & Medicinal Chemistry Letters·J YinC Pidgeon
May 13, 1999·Cell Research·K Sharma, Y Shi
Jun 15, 1999·Trends in Biochemical Sciences·D K Perry, Y A Hannun
Jun 15, 1999·Trends in Biochemical Sciences·K Hofmann, V M Dixit
Apr 27, 2000·The Journal of Biological Chemistry·S El BawabY A Hannun
Jun 8, 2000·The Journal of Organic Chemistry·H AzumaK Ogino
Sep 23, 2000·Chemistry and Physics of Lipids·A Nilsson, R D Duan

❮ Previous
Next ❯

Citations

Jun 25, 2003·Protein Expression and Purification·Willem F NieuwenhuizenFriedrich Götz
May 5, 2006·Organic Letters·Colin G FergusonGlenn D Prestwich
Oct 13, 2009·Journal of Lipid Research·Massimo AureliSandro Sonnino
Sep 20, 2012·Bioorganic & Medicinal Chemistry·Krishna P BhabakChristoph Arenz
Jan 26, 2007·ChemMedChem·Antonio DelgadoGemma Fabriás
Mar 17, 2007·Chembiochem : a European Journal of Chemical Biology·Carmen BediaGemma Fabriàs
Jul 30, 2015·Chemistry and Physics of Lipids·Essa M Saied, Christoph Arenz
Jun 4, 2013·Chembiochem : a European Journal of Chemical Biology·Krishna P BhabakChristoph Arenz
Mar 31, 2007·Skin Pharmacology and Physiology·E HoubenV Rogiers
Aug 20, 2015·Journal of Lipid Research·Mireia CasasampereGemma Fabrias
Sep 10, 2019·Frontiers in Cell and Developmental Biology·Monica Rolando, Carmen Buchrieser
Jul 7, 2018·Chemical Reviews·Carmanah D HunterChristopher W Cairo
Jun 15, 2006·Chemical Reviews·Stéphane GuillarmeArnaud Haudrechy
Mar 3, 2010·Progress in Lipid Research·Patricia GangoitiAntonio Gómez-Muñoz

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.