ELECTROPHORETIC, SPECTRAL, CATALYTIC AND IMMUNOCHEMICAL PROPERTIES OF HIGHLY PURIFIED CYTOCHROME-P-450 FROM SHEEP LUNG

Date

1990-01-01

Author

Adalı, Orhan

Metadata

Show full item record

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Item Usage Stats

122
views

0
downloads

1. 1. Cytochrome P-450LgM2 was purified from sheep lung microsomes in the presence of detergents, Emulgen 913 and cholate. 2. 2. The purification procedure involved the chromatography of the detergent solubilized microsomes on DEAE-cellulose and hydroxylapatite. 3. 3. Cytochrome P-450LgM2 was further purified on second DEAE-cellulose and hydroxylapatite columns. 4. 4. The specific content of the highly purified P-450LgM2 was 16–18 nmol P-450/mg protein and purified 164-fold. 5. 5. The yield was 16% of the initial content in microsomes. 6. 6. The SDS-polyacrylamide slab gel electrophoresis (PAGE) of the purified lung cytochrome P-450LgM2 showed one protein band having the monomer molecular weight of 49,500. 7. 7. The absolute CO-difference spectrum of dithionate-reduced P-450LgM2 gave a peak at 451 nm. 8. 8. When sheep lung cytochrome P-450LgM2 and P-450LM2 purified from liver of phenobarbital (PB)-induced rabbit were subjected to Western Blotting and visualized immunochemically with antiP-450LM2, they showed identical mobilities. 9. 9. P-450LgM2 was found to be very active in N-demethylation of benzphetamine in a reconstituted system containing purified sheep lung reductase and synthetic lipid. 10. 10. Turnover numbers (min−1) for benzphetamine, aniline, ethylmorphine and p-nitrophenol were determined to be 273, 1.2, 15.5 and 1.05, respectively, in a reconstituted microsomal lung monooxygenase system. 11. 11. Spectral, electrophoretic, biocatalytic and immunochemical properties of sheep lung P-450LgM2 were found to be similar to those of P-450 isozyme 2, purified from PB-treated rabbit liver and of rabbit lung microsomes.

Subject Keywords

Biochemistry

URI

https://hdl.handle.net/11511/39472

Journal

INTERNATIONAL JOURNAL OF BIOCHEMISTRY

DOI

https://doi.org/10.1016/0020-711x(90)90234-t

Collections

Department of Biology, Article

Suggestions

OpenMETU
Core

PURIFICATION OF NADH-CYTOCHROME-B5 REDUCTASE FROM SHEEP LUNG AND ITS ELECTROPHORETIC, SPECTRAL AND SOME OTHER PROPERTIES Güray, Nülüfer Tülün (Elsevier BV, 1990-01-01) 1. 1. NADH-cytochrome b5 reductase was purified from sheep lung microsomes in the presence of non-ionic and ionic detergents. Emulgen 913 and cholate, respectively. 2. 2. The purification procedure involved the ion-exchange chromatography of the detergent solubilized microsomes on DEAE-cellulose. 3. 3. Further purification and concentration of lung reductase was carried out with a second DEAE-cellulose column followed by the affinity column chromatography of partially purified reductase on 5'-ADP-agarose ...
SOLUBILIZATION AND PARTIAL-PURIFICATION OF 2 FORMS OF CYTOCHROME-P-450 FROM TROUT LIVER-MICROSOMES ARINC, E; Adalı, Orhan (Elsevier BV, 1983-01-01) 1. 1. Liver microsomal cytochrome P-450 content, NADPH-cytochrome reductase, aniline 4-hydroxylase and ethylmorphine activities of rainbow trout, Salmo gairdneri, were found to be 0.16 () nmol P-450/mg protein, 38 () units/mg protein, 0.04 () nmol protein and 0.174 ( ) nmol formaldehyde/min/mg protein, respectively. 2. 2. Trout liver cytochrome P-450 was solubilized by treatment of microsomes with sodium cholate. Chromatography on DEAE-cellulose column yielded two distinct cytochrome P-450 f...
STIMULATORY EFFECTS OF LUNG CYTOCHROME B5 ON BENZPHETAMINE N-DEMETHYLATION IN A RECONSTITUTED SYSTEM CONTAINING LUNG CYTOCHROME P450LGM2 ARINC, E; PASHA, RPK; Adalı, Orhan; BASARAN, N (Elsevier BV, 1994-08-01) 1. Cytochrome b 5 was partially purified from sheep lung microsomes in the presence of detergents Emulgen 913 and cholate by three consecutive DEAE-cellulose and Sephadex G-100 gel filtration chromatographies.
Synthesis of diols using the hypervalent iodine(III) reagent, phenyliodine(III) bis(trifluoroacetate) Celik, Murat; Alp, Cemalettin; Coşkun, Betül; Gultekin, M. Serdar; Balcı, Metin (2006-05-29) 1.2- and 1.3-Bis(trifluoroacetoxy) alcohols are easily obtained from the one-pot reaction of alkenes with phenyliodine(III) bis(trifluoroacetate) (PIFA) in the absence of any additive or catalyst. The products were converted into the corresponding diols by ammonolysis. The use of bicyclic alkenes has shown that rearranged 1,3-diacetoxy alcohols are mostly formed as the major products.
Crosslinked DADMAC polymers as cationic super absorbents Korpe, Sinan; Erdogan, Baris; Bayram, Göknur; Özgen, Serkan; Uludağ, Yusuf; Bicak, Niyazi (Elsevier BV, 2009-09-01) Loosely crosslinked cationic polyelectrolytes (polyquats) with super-water-sorption abilities are presented, for the first time. Hydrogels obtained by radically initiated copolymerization of N,N-diallyl, N,N-dimethyl ammonium chloride (DADMAC) with N-vinyl 2-pyrrolidone (NVP) in presence of N,N,N',N'-tetraallyl piperazinium dichloride (TAP) as crosslinker showed reasonably high equilibrium swelling ratios, as high as those for poly(acrylic acid)-based super absorbents.

Citation Formats

O. Adalı, “ELECTROPHORETIC, SPECTRAL, CATALYTIC AND IMMUNOCHEMICAL PROPERTIES OF HIGHLY PURIFIED CYTOCHROME-P-450 FROM SHEEP LUNG,” INTERNATIONAL JOURNAL OF BIOCHEMISTRY, pp. 1433–1432, 1990, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/39472.