Analytik plasmalogenhaltiger Phosphatide aus Herzgewebe mittels extracellulärer Phospholipase A<sub>2</sub>

Stoll, Ulrich

doi:10.1002/lipi.19960980108

Cited by 7 publications

(1 citation statement)

References 5 publications

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“…In fact snake venom sPLA 2 s were employed to analyse the position of fatty acids in phospholipids from guina pig and pic cardiac membranes [6]. Moreover, porcine PLA 2 (PPLA2) was used in the examination of the structural and functional changes of egg yolk low density lipoproteins (LDL) by modifying its phospholipids [7].…”

Section: Introductionmentioning

confidence: 99%

Proteolytic cleavage of stingray phospholipase A2: Isolation and biochemical characterization of an active N-terminal form

Bacha

Mejdoub

2011

Lipids Health Dis

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BackgroundMammalian GIB-PLA2 are well characterized. In contrast, much less is known about aquatic ones. The aquatic world contains a wide variety of living species and, hence represents a great potential for discovering new lipolytic enzymes. The aim of this study was to check some biochemical and structural properties of a marine stingray phospholipase A2 (SPLA2).ResultsThe effect of some proteolytic enzymes on SPLA2 was checked. Chymotrypsin and trypsin were able to hydrolyze SPLA2 in different ways. In both cases, only N-terminal fragments were accumulated during the hydrolysis, whereas no C-terminal fragment was obtained in either case. Tryptic and chymotryptic attack generated 13 kDa and 12 kDa forms of SPLA2, respectively. Interestingly, the SPLA2 13 kDa form was inactive, whereas the SPLA2 12 kDa form conserved almost its full phospholipase activity. In the absence of bile slats both native and 12kDa SPLA2 failed to catalyse the hydrolysis of PC emulsion. When bile salts were pre-incubated with the substrate, the native kinetic protein remained linear for more than 25 min, whereas the 12 kDa form activity was found to decrease rapidly. Furthermore, The SPLA2 activity was dependent on Ca2+; other cations (Mg2+, Mn2+, Cd2+ and Zn2+) reduced the enzymatic activity notably, suggesting that the arrangement of the catalytic site presents an exclusive structure for Ca2+.ConclusionsAlthough marine and mammal pancreatic PLA2 share a high amino acid sequence homology, polyclonal antibodies directed against SPLA2 failed to recognize mammal PLA2 like the dromedary pancreatic one. Further investigations are needed to identify key residues involved in substrate recognition responsible for biochemical differences between the 2 classes of phospholipases.

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Section: Introductionmentioning

confidence: 99%

Proteolytic cleavage of stingray phospholipase A2: Isolation and biochemical characterization of an active N-terminal form

Bacha

Mejdoub

2011

Lipids Health Dis

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Identifizierung von molekularen Phospholipid‐Species aus Schweineherz durch Kombination chromatographischer Techniken

Stoll

1996

Fett/Lipid

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over (Deutsc h land) Fur Herrn Dr. W. Umbach, Prasident der DGF, zu seinem 60. Geburtstag Die Phospholipide Phosphatidylcholin (PC) und Phosphatidylethanolamin (PE) aus subcellularen Systemen des Schweineherzens wurden nach Phospholipase C-Spaltung und Acetylierung mittels Dunnschichtchromatographie (DC) in die Subklassen Diacyl-, Alkylacylund Alkenylacyl-Diglyceride aufgetrennt. Das Diglyceridmuster jeder Subfraktion von PC bzw. PE wurde mittels Gaschromatographie (GC) auf einer TAP-Kapillarsaule bestimmt. Durch Acetylierung der Diradyl-Species konnten DC und G C ohne weitere Modifizierung kombiniert werden. Zur Identifizierung der Diradyl-Species wurde auch die Subfraktionierung auf Kieselgel-AgNOj-Schicht und das FettsaureIFettaldehydmuster nach Methylierung herangezogen. AuBerdem envies sich die Hydrierung der acetylierten Diglyceride zur Charakterisierung von PC und PE als giinstig und informativ. Die Subfraktionen innerhalb eines Phosphatids zeigen sehr ahnliche Profile fur die Diradyl-Species. Zwischen den Phosphatiden PC und PE sind jedoch deutliche Unterschiede im Muster zu erkennen.In PE sind die Species c&20:1 und solche mit Iangerkettigen Fettsauren in sn-2-Position sehr vie1 haufiger als bei PC. Die Fraktionierung in Subklassen ist von erheblicher Bedeutung, da sich Diacyl-Species sehr unterschiedlich zu Alkenylacyl-Species irn Stoffwechsel verhalten. Diradyl-Glyceride lassen sich auf einer TAP-Saule auch ohne Subfraktionierung mittels DC trennen, wegen der drei Subklassen pro Phosphatid im Herzgewebe ist diese Methode jedoch besonders angebracht. Identification of Molecular Species of Phospholipids from PigHeart by Combining Chromatographic Techniques. Glycerophospholipids from subcellular organelles and membranes from pig heart were divided into subclasses. The molecular species within the main subclasses were analyzed. The phosphorus moiety had been removed by phospholipase-C and the acetylated molecules were separated in diacyl-, alkylacyl-and alkenylacyl-glycerols by thin-layer chromatography (TLC). For further identification the subclasses from the first fractionation by TLC were separated by silver ion chromatography according to their polarity. A small portion of the acetylated diradyl-species was transesterified so that the fatty acid composition could be assessed as methyl esters. The acetylated molecular species derived from glycerophospholipids were chromatographed on WCOT fused silica columns with TAP as stationary phase according to molecular weight and unsaturation. Gaschromatography (GC) of fatty acid methyl esters and dimethylacetals from plasmalogens was executed on DEGS-columns or on WCOT fused silica columns with CP-Sil 88 as stationary phase. The distribution of molecular species within a phospholipid class (PC or PE) was rather similar for the diacyl-and the alkenylacyl molecules. But the latter are more volatile. In the case of phosphatidylcholine the composition OfC16-C18-I; CI6-CIB 2 and c18-Cl8 2 was predominant for diacyl-as well as alkenylacyl-species. In the case of phosp...

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Antibacterial, Anti-Chlamydial, and Cytotoxic Activities of a Marine Snail (Hexaplex trunculus) Phospholipase A2: an In Vitro Study

Zarai

Gharsallah

Hammami

et al. 2012

Appl Biochem Biotechnol

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In order to report pharmacological characterization of marine snail (Hexaplex trunculus) hepatopancreatic phospholipase A(2) (mSDPLA(2)), we have talked for the first time the antimicrobial activity against different pathogenic bacterial strains, anti-chlamydial activity as well as its cytotoxic activity against McCoy cell lines. mSDPLA(2), showed a high level of activity towards Gram-positive bacteria as Staphylococcus aureus and Staphylococcus epidermidis. Whereas Gram-negative bacteria, unfortunately, exhibited a higher resistance, mSDPLA(2) was also found to have a strong cytotoxic activity, causing significant morphological alterations of the McCoy cell lines surfaces and to be a hinder to the proliferation. Moreover, mSDPLA(2) proved to have a very potent anti-chlamydial activity. Over 95 % inhibition of chlamydial inclusions were obtained at a concentration of 10 μg/ml of mSDPLA(2) after 24 h postinfection. Interestingly, at a concentration of 10 μg/ml of mSDPLA(2), the proliferation of McCoy cells was not affected. Approximately 50 % inhibition of cell growth was obtained with a concentration of 37 μg/mL of mSDPLA(2). mSDPLA(2) could be considered as an excellent candidate for the development of a new anti-infective agent. This enzyme showed significant antimicrobial activities.

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Analytik plasmalogenhaltiger Phosphatide aus Herzgewebe mittels extracellulärer Phospholipase A₂

Cited by 7 publications

References 5 publications

Proteolytic cleavage of stingray phospholipase A2: Isolation and biochemical characterization of an active N-terminal form

Proteolytic cleavage of stingray phospholipase A2: Isolation and biochemical characterization of an active N-terminal form

Identifizierung von molekularen Phospholipid‐Species aus Schweineherz durch Kombination chromatographischer Techniken

Antibacterial, Anti-Chlamydial, and Cytotoxic Activities of a Marine Snail (Hexaplex trunculus) Phospholipase A2: an In Vitro Study

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Analytik plasmalogenhaltiger Phosphatide aus Herzgewebe mittels extracellulärer Phospholipase A2

Cited by 7 publications

References 5 publications

Proteolytic cleavage of stingray phospholipase A2: Isolation and biochemical characterization of an active N-terminal form

Proteolytic cleavage of stingray phospholipase A2: Isolation and biochemical characterization of an active N-terminal form

Identifizierung von molekularen Phospholipid‐Species aus Schweineherz durch Kombination chromatographischer Techniken

Antibacterial, Anti-Chlamydial, and Cytotoxic Activities of a Marine Snail (Hexaplex trunculus) Phospholipase A2: an In Vitro Study

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Analytik plasmalogenhaltiger Phosphatide aus Herzgewebe mittels extracellulärer Phospholipase A₂