Kaya Lutter

Heinrich Heine University Düsseldorf

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Wilhelm Stahl

Heinrich Heine University Düsseldorf

Tanja Wagener

Heinrich Heine University Düsseldorf

Sebastian Kock

Technische Universität Braunschweig

Hans‐Dieter Martin

Heinrich Heine University Düsseldorf

Helmut Görner

Max Planck Institute for Chemical Energy Conversion

Susanne Frixel

Max Planck Institute for Dynamics of Complex Technical Systems

Wolfgang Schräder

Max-Planck-Institut für Kohlenforschung

Roger Scherrers

Heinrich Heine University Düsseldorf

Klaus Schaper

Heinrich Heine University Düsseldorf

Hansgeorg Ernst

BASF (Germany)

Cooperative Institutions

Heinrich Heine University Düsseldorf

Consejo Superior de Investigaciones Científicas

University of Konstanz

Max Planck Society

Centre National de la Recherche Scientifique

Düsseldorf University Hospital

University of Münster

Centre de Génétique Moléculaire

GTx (United States)

Leibniz University Hannover

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Molecular Identification of a SNAP-25-Like SNARE Protein inParamecium

Eukaryotic Cell (2008)

Christina Schilde Kaya Lutter Roland Kissmehl Helmut Plattner

Using database searches of the completed Paramecium tetraurelia macronuclear genome with the metazoan SNAP-25 homologues, we identified a single 21-kDa Qb/c-SNARE in this ciliated protozoan, named P. tetraurelia SNAP (PtSNAP), containing the characteristic dual heptad repeat SNARE motifs of SNAP-25. The presence of only a single Qb/c class SNARE in P. tetraurelia is surprising in view of the multiple genome duplications and the high number of SNAREs found in other classes of this organism. As inferred from the subcellular localization of a green fluorescent protein (GFP) fusion construct, the protein is localized on a variety of intracellular membranes, and there is a large soluble pool of PtSNAP. Similarly, the PtSNAP that is detected with a specific antibody in fixed cells is associated with a number of intracellular membrane structures, including food vacuoles, the contractile vacuole system, and the sites of constitutive endo- and exocytosis. Surprisingly, using gene silencing, we could not assign a role to PtSNAP in the stimulated exocytosis of dense core vesicles (trichocysts), but we found an increased number of food vacuoles in PtSNAP-silenced cells. In conclusion, we identify PtSNAP as a Paramecium homologue of metazoan SNAP-25 that shows several divergent features, like resistance to cleavage by botulinum neurotoxins.

Snap

Identification

SNARE complex

Vesicular Transport Proteins

10.1128/ec.00012-08

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Citations (26)

3,3′‐Dihydroxyisorenieratene, a Natural Carotenoid with Superior Antioxidant and Photoprotective Properties

Angewandte Chemie International Edition (2008)

Hans‐Dieter Martin Sebastian Kock Roger Scherrers Kaya Lutter Tanja Wagener

The color of red smear cheeses arises from naturally occurring carotenoids like 3,3′-dihydroxyisorenieratene (1). This compound and its oxidation product 2, which have unusual phenolic and quinoid end groups, respectively, were synthesized. The antioxidant and photoprotective properties of a series of carotenoids were tested with eight different model systems, and 1 proved to be superior to other carotenoids. Detailed facts of importance to specialist readers are published as "Supporting Information". Such documents are peer-reviewed, but not copy-edited or typeset. They are made available as submitted by the authors. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

Natural product

10.1002/anie.200803668

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Molecular Identification of 26 Syntaxin Genes and their Assignment to the Different Trafficking Pathways in Paramecium

Traffic (2007)

Roland Kissmehl Christina Schilde Thomas Wassmer Carsten Friedrich Danzer Kathrin Nuehse

SNARE proteins have been classified as vesicular (v)‐ and target (t)‐SNAREs and play a central role in the various membrane interactions in eukaryotic cells. Based on the Paramecium genome project, we have identified a multigene family of at least 26 members encoding the t‐SNARE syntaxin ( Pt Syx) that can be grouped into 15 subfamilies. Paramecium syntaxins match the classical build‐up of syntaxins, being ‘tail‐anchored’ membrane proteins with an N‐terminal cytoplasmic domain and a membrane‐bound single C‐terminal hydrophobic domain. The membrane anchor is preceded by a conserved SNARE domain of ∼60 amino acids that is supposed to participate in SNARE complex assembly. In a phylogenetic analysis, most of the Paramecium syntaxin genes were found to cluster in groups together with those from other organisms in a pathway‐specific manner, allowing an assignment to different compartments in a homology‐dependent way. However, some of them seem to have no counterparts in metazoans. In another approach, we fused one representative member of each of the syntaxin isoforms to green fluorescent protein and assessed the in vivo localization, which was further supported by immunolocalization of some syntaxins. This allowed us to assign syntaxins to all important trafficking pathways in Paramecium.

Syntaxin 3

SNARE complex

10.1111/j.1600-0854.2007.00544.x

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Citations (45)

3,3′‐Dihydroxyisorenieratene prevents UV‐induced formation of reactive oxygen species and the release of protein‐bound zinc ions in human skin fibroblasts

Molecular Nutrition & Food Research (2009)

Kaya Lutter Silke De Spirt Sebastian Kock Klaus‐Dietrich Kröncke Hans‐Dieter Martin

3,3'-Dihydroxyisorenieratene (DHIR) is a structurally unusual carotenoid exhibiting bifunctional antioxidant properties. It is synthesized by Brevibacterium linens, used in dairy industry for the production of red smear cheeses. The compound protects cellular structures against photo-oxidative damage and inhibits the UV-dependent formation of thymidine dimers. Here we show that DHIR prevents a UV-induced intracellular release of zinc ions from proteins in human dermal fibroblasts. The effect is correlated with a decreased formation of intracellular reactive oxygen species. In contrast, zinc release from cellular proteins induced by hyperthermia is not affected by pretreatment of cells with the antioxidant DHIR. It is suggested that the intracellular zinc release upon UV irradiation is due to oxidative modifications of the zinc ligands in proteins (e.g. cysteine) and that protection by DHIR is due to intracellular scavenging of reactive oxygen species generated in photo-oxidation.

10.1002/mnfr.200900044

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Citations (9)

Titelbild: 3,3′‐Dihydroxyisorenieratin, ein natürliches Carotinoid mit überlegenen antioxidativen und photoprotektiven Eigenschaften (Angew. Chem. 2/2009)

Angewandte Chemie (2008)

Hans‐Dieter Martin Sebastian Kock Roger Scherrers Kaya Lutter Tanja Wagener

10.1002/ange.200890330

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Carotenoids in Photooxidative Stress

Current Nutrition & Food Science (2010)

Silke De Spirt Kaya Lutter Wilhelm Stahl

Carotenoids are secondary plant constituents and more than 700 different compounds have been identified. They are synthesized by plants, where they serve as colorants for fruits and leaves, bacteria, fungi and algae. In nature carotenoids are important biological compounds due to their provitamin A activity, antioxidant properties and accessory functions in the light harvesting system of plants. Considerable amounts of carotenoids are ingested with the diet and accumulate in the human organism. α- and β- carotene, β- cryptoxanthin, lutein, zeaxanthin and lycopene are the major carotenoids in human blood and tissues. Based on their structural features carotenoids are suitable compounds for photoprotection in humans. They may act as ultraviolet (UV) absorbers and dietary antioxidants capable of scavenging reactive intermediates generated under the condition of photooxidative stress. Photooxidative stress is involved in processes of photoageing, photocarcinogenesis and plays a major role in the pathogenesis of photodermatoses. Intervention studies with β- carotene and lycopene supplements or diets rich in those carotenoids have shown that these molecules contribute to systemic photoprotection ameliorating UVinduced erythema. In- vitro data provide evidence that also other carotenoids are efficient photoprotectors, for example lutein and the structurally unusual phenolic polyene 3,3- dihydroxyisorenieratene. Keywords: Carotenoids, UV, photoprotection, skin, DHIR

Photoprotection

Phytoene

Carotene

10.2174/157340110790909572

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Citations (15)

Untersuchungen zur photoprotektiven Wirkung von 3,3'-Dihydroxyisorenieratin und Vitamin E Derivaten

Kaya Lutter

Die menschliche Haut ist neben dem Auge das einzige lichtexponierte Organ. Ohne ausreichenden Schutz, u.a. durch korpereigene Substanzen oder durch die topische Anwendung von Sonnenschutzmitteln, fuhrt ubermasige Sonnenstrahlung, insbesondere Exposition gegenuber energiereicher UV-Strahlung, zu Schadigungen der Haut. Dabei handelt es sich um direkte entstehende Schaden, bei denen UV-Strahlen unmittelbar mit biologisch relevanten Molekulen interagieren (z.B. der DNA), oder um indirekte Schaden, welche uber sekundare, UV-induzierte Reaktionsprodukte vermittelt werden (z.B. Photooxidation). Zu den sekundaren Intermediaten zahlen die reaktiven Sauerstoffverbindungen (ROS), sowohl freie Radikale wie Superoxidradikalanionen, Peroxylradikale und Hydroxylradikale, als auch nichtradikalische Verbindungen wie Singulettsauerstoff oder Wasserstoffperoxid. Als photooxidativer Stress wird ein Ungleichgewicht bezeichnet bei dem lichtinduzierte oxidative Schaden nicht vollstandig von den Schutzsystemen verhindert werden und in dessen Verlauf ROS zellulare Strukturen wie DNA, Proteine und Lipide oxidieren. Photooxidativer Stress spielt eine wichtige Rolle in der Pathogenese von Hautkrebserkrankungen, bei vorzeitiger Hautalterung, Veranderung des cutanen Immunsystems und bei der Entwicklung von Photodermatosen. Antioxidative Mikronahrstoffe wie die Vitamine E und C sowie die Substanzgruppe der Carotinoide werden zur endogenen Photoprotektion beim Menschen eingesetzt. Als orale Sonnenschutzmittel tragen sie zur antioxidativen Verteidigung bei. DHIR ist ein phenolisches Carotinoid und kommt u.a. im Brevibacterium linens vor, welches in der Nahrungsmittelindustrie Verwendung findet. In der vorliegenden Arbeit wurden die UV-protektiven Eigenschaften von DHIR in Zellkulturexperimenten an humanen Hautfibroblasten im Vergleich zu Lutein untersucht. Keine der Verbindungen war mit oder ohne UV-Bestrahlung zelltoxisch (Sulforhodamin B Test). Als zellulare Antwort auf UV-Bestrahlung wurde die Expression des Enzyms Hamoxygenase 1 (HO-1) anhand der Proteinmenge mittels Western Blot untersucht. Nach UV-A induzierter Photooxidation ist die Expression der HO-1 erhoht. Im Vergleich zur bestrahlten Kontrolle (20 J/cm²) war die Expression von HO-1 um 30% geringer, wenn die Zellen uber 24 h vor Bestrahlung mit 1,5 µM DHIR behandelt wurden. Bei gleicher Vorbehandlung mit Lutein wurden keine Effekte beobachtet. DHIR verhindert die Freisetzung von Zn-Ionen aus Proteinkomplexen, was auf einen Schutz der Thiolliganden vor Oxidation zuruckgefuhrt wird. Die Bildung von Cyclobutanpyrimidindimeren ist eine Hauptschadigung der DNA nach UV-B Exposition. In den Fibroblasten konnten nach Bestrahlung mit UV-B (300 mJ/cm²) Thymidindimere mit spezifischen Antikorpern nachgewiesen werden. Waren die Zellen 24 h mit DHIR vorbehandelt bilden sich weniger Thymidindimere. Lutein hatte keinen Effekt. Da die Bildung von Cyclobutanpyrimidindimeren eine photochemische Reaktion der DNA Basen ist und nicht mit photooxidativen Prozessen zusammenhangt sind hier wahrscheinlich UV-B absorbierende Eigenschaften fur den Schutzeffekt verantwortlich. Das naturliche Carotinoid DHIR verfugt uber ausergewohnliche antioxidative und photoprotektive Eigenschaften. Ursache dafur ist die besondere Struktur des Carotinoids, welches aufgrund polyenischer und phenolischer Strukturelemente als bifunktioneller Radikalfanger wirken kann und gleichzeitig UV-Licht absorbiert. In der Arbeit untersuchte synthetische Vitamin E Derivate erwiesen sich ebenfalls als photoprotektiv, was mit einer Verringerung der HO-1 Expression nach UVA-Bestrahlung belegt wurde. Keine der Substanzen war aber dem naturliche Vitamer in der Wirkung uberlegen.

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Cover Picture: 3,3′‐Dihydroxyisorenieratene, a Natural Carotenoid with Superior Antioxidant and Photoprotective Properties (Angew. Chem. Int. Ed. 2/2009)

Angewandte Chemie International Edition (2008)

Hans‐Dieter Martin Sebastian Kock Roger Scherrers Kaya Lutter Tanja Wagener

The red smear cheeses shown on the cover obtain their color from bacteria such as Brevibacterium linens, which contain highly effective antioxidant and photoprotective phenolic carotenoids such as 3,3′-dihydroxyisorenieratene (DHIR). In their Communication on page 400 ff., H.-D. Martin, W. Stahl, and co-workers describe the synthesis of DHIR and their investigation of its antioxidative and photoprotective properties. Its quinoid oxidation product is blue in color.

Brevibacterium

Photoprotection

10.1002/anie.200890281

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3,3′‐Dihydroxyisorenieratin, ein natürliches Carotinoid mit überlegenen antioxidativen und photoprotektiven Eigenschaften

Angewandte Chemie (2008)

Hans‐Dieter Martin Sebastian Kock Roger Scherrers Kaya Lutter Tanja Wagener

Eine farbgebende Komponente von Rotschimmelkäse ist das bakterielle Carotinoid 3,3′-Dihydroxyisorenieratin (1). Nun ist es gelungen, 1 sowie dessen Oxidationsprodukt 2 zu synthetisieren; beide Verbindungen haben außergewöhnliche Strukturen mit phenolischen bzw. chinoiden Endgruppen. Anhand von acht Modellsystemen wurden die antioxidativen und photoprotektiven Eigenschaften untersucht, wobei sich 1 als anderen Carotinoiden überlegen erwies.

10.1002/ange.200803668

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Citations (1)

Photoprotective effects of aromatic carotenoids in human dermal fibroblasts (hdF)

Acta Biologica Cracoviensia. Series Zoologia (2011)

Sarah Wagener Kaya Lutter Silke De Spirt Wilhelm Stahl

Dermal fibroblast

Human skin

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