Purification of CYP2B-like protein from feral leaping mullet (Liza saliens) liver microsomes and its biocatalytic, molecular, and immunological characterization

Arinc, Emel
In this study, CYP2B-immunoreactive protein was purified to electrophoretic homogeneity from the liver microsomes of leaping mullet. The purified cytochrome P450 (CYP) gave a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis having a M, of 49,300 Da. Absolute absorption spectrum of the purified CYP showed a maximum at 417 nm and CO-difference spectrum of dithionite-reduced cytochrome P450 gave a peak at 450 nm. The purified CYP was found to be active in N-demethylation of benzphetamine, erythromycin, and ethylmorphine, and O-dealkylation of pentoxyresorufin in the reconstituted system. However, it was unable to catalyze O-dealkylation of ethoxyresorufin, methoxyresorufin, benzyloxyresorufin, and hydroxylation of lauric acid and aniline. The purified CYP showed strong cross-reactivity with anti-sheep lung CYP2B, a homologue of CYP2B4. N-terminal amino acid sequence of the mullet P450 had the highest degree of homology with CYP2Bs among the known CYPs. Spectral, electrophoretic, immunochemical, N-terminal amino acid sequence, and biocatalytic properties of the purified CYP are most similar to those of mammalian cytochrome P4502B. All these data indicate that the purified CYP is certainly 2B-like. In this study, we not only purified biocatalytically active CYP2B-like protein from fish, but also demonstrated detailed functional properties of CYP2B-like protein for the first time. (C) 2008 Wiley Periodicals, Inc.


Biochemical characteristics of purified beef liver NADPH-cytochrome P450 reductase
Arinc, E; Celik, H (Wiley, 2002-01-01)
NADPH-cytochrome P450 reductase, an obligatory component of the cytochrome P450 dependent monooxygenase system, was purified to electrophoretic homogeneity from beef liver microsomes. The purification procedure involved the ion exchange chromatography of the detergent-solubilized microsomes on first and second DEAE-cellulose columns, followed by 2',5'-ADP Sepharose affinity chromatography. Further concentration of the enzyme and removal of Emulgen 913 and 2'-AMP were accomplished on the final hydroxylapatit...
Purification and Characterization of Cytochrome P450 Reductase from Liver Microsomes of Feral Leaping Mullet (Liza saliens)
Sen, Alaattin; Arinc, Emel (Wiley, 1998-01-01)
NADPH-cytochrome P450 reductase was purified to electrophoretic homogeneity from detergent-solubilized liver microsomes from the leaping mullet (Liza saliens). The purified reductase was characterized with respect to spectral, electrophoretic, and biocatalytic properties. In addition, effects of pH, ionic strength, and the substrate concentration on the NADPH-dependent cytochrome c reductase activity of the purified fish liver cytochrome P450 reductase were studied. Cytochrome P450 reductase was purified 43...
Characterization of proteome alterations in Phanerochaete chrysosporium in response to lead exposure
Yildirim, Volkan; ÖZCAN, Servet; Becher, Doerte; Buettner, Knut; Hecker, Michael; Özcengiz, Gülay (Springer Science and Business Media LLC, 2011-03-09)
Background: Total soluble proteome alterations of white rot fungus Phanerochaete chrysosporium in response to different doses (25, 50 and 100 mu M) of Pb (II) were characterized by 2DE in combination with MALDI-TOF-MS.
An alternative supporting electrolyte for enzyme immobilization in conducting polymers
Kiralp, Senem; Balik, Balam; KARATAŞ, SEVİM; Toppare, Levent Kamil; Gungorb, Atilla (Elsevier BV, 2008-03-01)
In this study an alternative supporting electrolyte was used in enzyme immobilization. Invertase was studied to observe the effect of the supporting electrolyte. Sulfonated poly(arylene ether sulfone) was used as the supporting electrolyte during the electrolysis of pyrrole. The results show that the polymeric supporting electrolyte can be used instead of sodium dodecyl sulfate.
Biomarkers of marine pollution observed in species of mullet living in two eastern Mediterranean harbours
Karakoc, FT; Hewer, A; Phillips, DH; Gaines, AF; Yuregir, G (Informa UK Limited, 1997-09-01)
The activities of enzymes associated with xenobiotic metabolism and/or oxidative processes, and the levels of aromatic-DNA adducts, have been determined in the livers of grey mullet (Oedalechilus labeo and Lisa ramada) living in two eastern Mediterranean harbours. Glutathione peroxidase (GSH-P) activity was 2.5 times higher (similar to 9 IU g(-1) liver) and glutathione reductase (GSSG-R) activity was twice as high (similar to 2.5 id g(-1) liver) in fish from the more polluted harbour at Mersin than in the h...
Citation Formats
A. BOZCAARMUTLU and E. Arinc, “Purification of CYP2B-like protein from feral leaping mullet (Liza saliens) liver microsomes and its biocatalytic, molecular, and immunological characterization,” JOURNAL OF BIOCHEMICAL AND MOLECULAR TOXICOLOGY, pp. 284–298, 2008, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/64491.