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Conversion of phenoloxidase and peroxidase indicators in individual haemocytes of Mytilus edulis specimens and isolation of phenoloxidase from haemocyte extract (1996)

Renwrantz, L. ; Schmalmack, W. ; Redel, R. ; [et al.]

Springer

Journal of comparative physiology 165 (1996), S. 647-658

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ISSN: 1432-136X

Keywords: Cell mobility ; Cytotoxic haemocytes ; Phenoloxidase ; Peroxidase indicators ; Mussel, Mytilus edulis

Source: Springer Online Journal Archives 1860-2000

Topics: Biology , Medicine

Notes: Abstract Haemocytes oxidized 3-amino-9-ethylcarbazole and other peroxidase indicators such as 3,3′-diaminobenzidine·4HCl, 3,3′,5,5′ tetramethylbenzidine·2HCl and 4-chloro-1-naphthol without addition of H2O2 indicating that the reaction was possibly not caused by a peroxidase. As these chromogens were also converted by a mushroom phenoloxidase in the absence of H2O2, cell smears were incubated with known substrates of phenoloxidases. One of these, l-dopa, caused strong melanin formation in several haemocytes and the reaction could be blocked by a variety of inhibitors including KCN, NaF, 1-phenyl-2-thiourea, cysteine, glutathione, ascorbic acid and HgCl2. The enzymatic activity was isolated using a concanavalin A column and separated into two fractions with an ion-exchange cartridge. The molecular weights of the glycoproteins were estimated to be 381±13.7 kDa and 316±11.1 kDa. After isoelectric focusing of a haemocyte extract and the two ion-exchange peaks, seven enzyme bands were detected with isoelectric points between pH 5.0 and 5.5. The isolated enzyme fractions both converted 3,3′,5,5′-tetramethylbenzidine·2HCl best at pH 5–6 and l-dopa at pH 7.0 without addition of H2O2. Heat-treated cells lost their enzymatic activity; however, a group of haemocytes still bound preoxidized 3-amino-9-ethylcarbazole (= AECox). Also, some of the phenoloxidase inhibitors mentioned above blocked this non-enzymatic staining reaction. About 30–57% of haemocytes from individual mussels were AECox-positive, whereas Mytilus specimens without phenoloxidase-containing cells often occurred. Haemocytes containing this enzyme exhibited a high mobility and a large percentage of them belonged to a cytotoxic cell population.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF00301133

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[Cu4(NPMe3)3(O2C—CH3)5] - ein tetramerer Phosphaniminato-Komplex von Kupfer(II). Synthese, Kristallstruktur, magnetisches Verhalten und EPR-Spektrum (1995)

Zu Köcker, R. Meyer ; Pebler, J. ; Friebel, C. ; [et al.]

Weinheim : Wiley-Blackwell

Zeitschrift für anorganische Chemie 621 (1995), S. 1311-1317

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ISSN: 0044-2313

Keywords: Phosphorane Iminato Complexes ; Copper(II) ; Synthesis ; Magnetic Behaviour ; EPR Spectrum ; Crystal Structure ; Chemistry ; Inorganic Chemistry

Source: Wiley InterScience Backfile Collection 1832-2000

Topics: Chemistry and Pharmacology

Description / Table of Contents: [Cu4(NPMe3)3(O2C—CH3)5] - a Tetrameric Phosphorane Iminato Complex of Copper(II). Synthesis, Crystal Structure, Magnetic Behaviour, and EPR SpectrumThe title compound and the corresponding benzoate complex [Cu4(NPMe3)3(O2C—C6H5)5] have been prepared by reactions of copper(II)acetate and copper(II)benzoate, respectively, with Me3SiNPMe3 in dichloromethane. Both complexes are characterized by IR spectroscopy. The acetate complex is additionally characterized by the measurement of the magnetic susceptibility, by its EPR spectrum, and by a crystal structure determination.[Cu4(NPMe3)3(O2C—CH3)5] · CH2Cl2: Space group I41/a, Z = 16, structure solution with 7 960 independent reflections, R = 0.044. Lattice dimensions at -70°C: a = b = 3 670.6; c = 1 091.9 pm. The structure consists of four Cu atoms which are arranged at the corners of a distorted tetrahedron with Cu…Cu distances between 290 and 318 pm. Three of the faces of the tetrahedron are linked by μ3-N atoms of the phosphorane iminato groups. Three of the acetate ligands form chelates, the other two are monofunctionally coordinated. Three of the copper atoms have a planar surrounding, the forth Cu atom has a (4 + 1) coordination.

Notes: Die Titelverbindung sowie der entsprechende Benzoat-Komplex [Cu4(NPMe3)3(O2C—C6H5)5] wurden durch Reaktion von Kupfer(II)acetat bzw. von Kupfer(II)benzoat mit Me3SiNPMe3 in Dichlormethan hergestellt und durch ihre IR-Spektren charakterisiert. Der Acetatkomplex wurde zusätzlich durch die Messung der magnetischen Suszeptibilität, des EPR-Spektrums und durch eine Kristallstrukturanalyse charakterisiert.[Cu4(NPMe3)3(O2C—CH3)5] · CH2Cl2: Raumgruppe I41/a, Z = 16, Strukturlösung mit 7 960 unabhängigen Reflexen, R = 0,044. Gitterkonstanten bei -70°C: a = b = 3 670,6; c = 1 091,9 pm. Die Struktur besteht aus vier Cu-Atomen, die ein verzerrtes Tetraeder bilden mit Cu…Cu-Abständen zwischen 290 und 318 pm. Drei der Tetraederflächen sind durch μ3-N-Atome dreier NPMe3--Gruppen überbrückt, drei der Acetatliganden sind chelatartig gebunden, während zwei Acetatgruppen monofunktionell koordinieren. Drei der Cu-Atome haben planare Umgebung, das vierte hat eine (4 + 1)-Koordination.

Additional Material: 5 Ill.