Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “CYTOCHROMES”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 973 records · Page 54Linked to original sources
PubMed1994

Interaction between cytochrome P450 2B1 and cytochrome bs: inhibition by synthetic peptides indicates a role for P450 residues Lys-122 and Arg-125.

Binding of cytochrome b5 to rat cytochrome P450 2B1 was inhibited (by 75%) by a synthetic peptide corresponding to P450 residues 116-134. The role of Lys-122 and Arg-125 were evaluated using peptides in which one or both of these basic residues were replaced with Glu. The Lys-122 substitution nearly abolished while the Arg-125 replacement decreased (by 20%) the inhibitory potential of the peptide. Substitution of both residues resulted in a peptide with no inhibitory activity. These results thus indicate a role for a specific P450 region as well as two basic residues within this region in the cytochrome P450-cytochrome b5 interaction.

Amino Acid Sequence↗
PubMed1995

Thermodynamic characterization of the interaction between cytochrome b5 and cytochrome c.

In recent years many studies have been directed toward the elucidation of the interaction mechanisms within protein-protein complexes. One of the best studies protein-protein complexes has been the cytochrome b5 and cytochrome c electron transfer pair. Thermodynamic information about the association process has been obtained through methods which indirectly measure the binding between the proteins. We report here the use of Isothermal Titration Calorimetry to characterize the association of Rat cytochrome b5 and Horse cytochrome c. The association is accompanied by an unfavorable enthalpy change (+1.0 +/-0.1 Kcal/mole) and a large stabilizing change in entropy (33.9 +/-0.6 eu).

Animals↗
PubMed1988

The measurement of cytochrome b559 in polymorphonuclear leukocytes and macrophages in the presence of hemoglobin or mitochondrial cytochromes.

A cyanide-insensitive b-type cytochrome (cytochrome b559) is involved in the respiratory burst of phagocytes. Spectroscopic measurement of its content in polymorphonuclear leukocytes or macrophages is complicated by signals arising from contaminating Hb or mitochondrial cytochromes upon reduction by dithionite. These interferences can be overcome by reducing the sample with dithionite, aerating it, and reducing it a second time with dithionite, in the presence of 10 microM cyanide. Cyanide suppresses the response of the interfering proteins to the second dithionite reduction. This modified method allows an accurate determination of the content of cytochrome b559 using smaller numbers of cells than previously feasible.

Animals↗
PubMed1990

Isolation of cytochrome P450 from hepatopancreas microsomes of the spiny lobster, Panulirus argus, and determination of catalytic activity with NADPH cytochrome P450 reductase from vertebrate liver.

One major form of cytochrome P450 has been isolated from the hepatopancreas of untreated spiny lobsters, Panulirus argus. This form, termed here D1, was purified to a specific content of 12.1 +/- 1.8 nmol/mg protein. Two minor forms, termed D2 and D3 were partially purified to 4.6 +/- 1.6 and 2.3 +/- 0.2 nmol P450/mg protein, respectively. No NADPH-cytochrome P450 reductase activity was detected in spiny lobster hepatopancreas microsomes and no purification of spiny lobster reductase was achieved in this study. Very low NADPH-cytochrome c reductase activity was found in hepatopancreas microsomes and also in cytosol. Indirect evidence suggested that proteolysis of spiny lobster P450 reductase during the preparation of hepatopancreas microsomes may in part account for the lack of detectable monooxygenase activity in hepatopancreas microsomes. The catalytic activities of the D1 or D2 forms of spiny lobster P450 were measured by mixing D1 or D2 with NADPH-cytochrome P450 reductase isolated from pig or rat liver microsomes. D2 was very efficient in demethylating benzphetamine, with a turnover number of 122 per minute, and D1 was an efficient catalyst of progesterone 16 alpha-hydroxylation, with a turnover number of 43 per minute. Other good substrates for D1 and D2 forms were aminopyrine, testosterone, benzo(a)pyrene, and 7-ethoxycoumarin. Little activity was found with methyl-, ethyl-, pentyl-, or benzyl-phenoxazone ethers as substrates. The profile of metabolites formed by D1 or D2 with benzo(a)pyrene as substrate were more similar to those formed with uninduced rat liver microsomes than to those formed by liver microsomes from uninduced flatfish species.

Amino Acids↗
PubMed1975

A kinetic completion of the cyclic photosynthetic electron pathway of Rhodopseudomonas sphaeroides: cytochrome b-cytochrome c2 oxidation-reduction.

In Rhodopseudomonas sphaeroides, following a single-turnover flash of light, cytochrome c2 is oxidized by reaction center bacteriochlorophyll, and a cytochrome b is reduced by the primary electron acceptor, probably via ubiquinone. In this report we show that, in the uncoupled state, the rate of re-oxidation of the cytochrome b is identical to the rate of reduction of the cytochrome c2, a kinetic completion of the cyclic photosynthetic electron transport system.

Antimycin A↗
PubMed1985

Binding of cytochrome c2 to the isolated reaction center of Rhodospirillum rubrum involves the "backside" of cytochrome c2.

Lys 109, Lys 112 and Glu 1 of cytochrome c2 from Rhodospirillum rubrum G-9 are about 4-fold less reactive towards acetic anhydride when cytochrome c2 is bound to the isolated photosynthetic reaction center from the same organism. The three shielded residues are clustered together on the "backside" of cytochrome c2. This contrasts with mitochondrial cytochrome c where "frontside" lysines are protected by different physiological electron transfer partners.

Acetylation↗
PubMed1991

Inducing effect of oxfendazole on cytochrome P450IA2 in rabbit liver. Consequences on cytochrome P450 dependent monooxygenases.

Male New Zealand rabbits were dosed with either 0.9, 4.5 or 22.5 mg/kg/day of oxfendazole by gastric intubation for 10 days. Oxfendazole administered at the therapeutic dose (4.5 mg/kg) and at the highest dose (22.5 mg/kg) increased 1.54- and 2.36-fold the total liver microsomal cytochrome P450 and more particularly the isoenzyme P450IA2 (95 and 184% increases) as demonstrated by western blotting. Increases in ethoxyresorufin O-deethylation and hydroxylations of benzopyrene and acetanilide occurred in livers of the same animals without any change in N-demethylation of aminopyrine, benzphetamine or erythromycin. Because of the unchanged level of mRNA specific to cytochrome P450IA2, as shown by northern blot analysis of poly mRNA, an enzyme stabilization rather than a transcriptional activation of IA2 genes should be involved in the P450IA2 regulation mechanisms. Oxfendazole bound strongly to cytochrome P450, giving rise to a type II spectrum, and inhibited noncompetitively the ethoxyresorufin O-deethylase and acetanilide hydroxylase activities, this confirmed that oxfendazole interacts only with the P450IA2 family. On the basis of a comparison of the enzymatic activities induced by various imidazole drugs, it was concluded that oxfendazole, like omeprazole and albendazole, behaved as a 3-methylcholanthrene-type inducer. These three benzimidazoles did not all belong to the same category of cytochrome P450 inducers as the antifungal drugs miconazole, clotrimazole and ketoconazole.

Animals↗
PubMed1983

Possible involvement of NADPH-cytochrome P450 reductase and cytochrome b5 on beta-ketostearoyl-CoA reduction in microsomal fatty acid chain elongation supported by NADPH.

The chain elongation products from [2-14C]malonyl-CoA and palmitoytl-CoA by rat liver microsomes initiated by NADPH were found mainly to be 18:1 and 18:0. When anti-NADPH-cytochrome P450 reductase or anti-cytochrome b5 immunoglobulin G were included in this system, not only was the overall chain elongation significantly suppressed, but also a new radioactive product, which was identified as 2-heptadecanone derived from an intermediate beta-ketostearate, is accumulated depending upon the amounts of IgG added. These results suggest that NADPH-cytochrome P450 reductase and cytochrome b5 participate in the conversion from beta-ketostearoyl-CoA to beta-hydroxystearoyl-CoA, which is the first reductive step of the microsomal chain elongating system.

Acyl Coenzyme A↗
PubMed1983

Thermodynamic investigations of cytochrome b5 unfolding. II. Detergent-solubilized cytochrome b5 in solution and in a reconstituted system with dimyristoyl phosphatidylcholine.

Thermal unfolding of the detergent-solubilized cytochrome b5 was investigated by scanning calorimetry. The protein shows different thermostability in the presence and absence of detergent, and it achieves the maximal transition temperature after incorporation into dimyristoyl phosphatidylcholine liposomes. However, transition temperature and Gibbs energy change at unfolding are still lower than that of the tryptic fragment of cytochrome b5 in aqueous solution. Cytochrome b5 undergoes in aqueous solution in the absence of detergent an irreversible, complicated transition, but it remains in the associated state after thermal denaturation. Half transition temperature, enthalpy and heat capacity changes of cytochrome b5 unfolding under various external conditions are reported and compared with the corresponding values of the tryptic fragment of the protein. The thermodynamic data and independent results are suitable for detailing a model proposed by Tanford (The Hydrophobic Effect (1980), pp. 205-211, John Wiley & Sons, New York) for the spatial arrangement of the protein within the membrane.

Animals↗
PubMed1996

Chromatographic separation and behavior of microsomal cytochrome P450 and cytochrome b5.

The methods used for separation of the multiple mammalian cytochrome P450 enzymes by liquid chromatography are reviewed. In addition to the chromatographic techniques, preparation and handling of samples and prefractionation procedures are considered. Conditions that affect stability and chromatographic resolution of cytochromes P450 are also discussed. Special emphasis is put on useful methods which are not routinely used for P450 separation, such as immobilized metal affinity or hydrophobic-interaction chromatography. Applications of low- and high-pressure methods with regard to preparative and analytical separations are compared. It is shown that high- and medium-pressure ion-exchange chromatography are suitable tools for separation of closely related P450 enzymes, especially when specific detection methods are available. In addition to fractionation of cytochromes P450, the isolation and chromatographic behavior of cytochrome b5 is discussed.

Animals↗
PubMed2005

A cytosolic cytochrome b5-like protein in yeast cell accelerating the electron transfer from NADPH to cytochrome c catalyzed by Old Yellow Enzyme.

A 410-nm absorbing species which enhanced the reduction rate of cytochrome c by Old Yellow Enzyme (OYE) with NADPH was found in Saccharomyces cerevisiae. It was solubilized together with OYE by the treatment of yeast cells with 10% ethyl acetate. The purified species showed visible absorption spectra in both oxidized and reduced forms, which were the same as those of the yeast microsomal cytochrome b5. At least 14 amino acid residues of the N-terminal region coincided with those of yeast microsomal b5, but the protein had a lower molecular weight determined to be 12,600 by SDS-PAGE and 9775 by mass spectrometry. The cytochrome b5-like protein enhanced the reduction rate of cytochrome c by OYE, and a plot of the reduction rates against its concentration showed a sigmoidal curve with an inflexion point at 6x10(-8) M of the protein.

Catalysis↗
PubMed1991

Some recent advances relating to prokaryotic cytochrome c reductases and cytochrome c oxidases.

Prokaryotic systems provide excellent experimental opportunities for exploring structure/function relationships for the complex, membrane-bound, multisubunit enzymes responsible for the reduction and subsequent oxidation of c-type cytochromes in respiratory or photosynthetic electron transport chains. Two points are made in this mini-review: (1) The eukaryotic and prokaryotic aa3-type cytochrome c oxidases are members of an apparently large superfamily of structurally related respiratory oxidases. This superfamily displays considerable variation in terms of the heme prosthetic groups (a or b) as well as the substrate oxidized (quinol or cytochrome c). The relationships among these enzymes help to facilitate explorations of how they work. (2) Molecular biology techniques can be used to generate intact, redox-active, water-soluble domains of membrane-bound subunits. These soluble domains can be used for detailed examination, including obtaining high resolution structure by NMR techniques or by X-ray crystallography. This approach is being used to study the soluble heme-binding domain of cytochrome c1 from the bc1 complex of Rhodobacter sphaeroides.

Cytochrome c Group↗
PubMed1997

Cytochrome P4501A1 and cytochrome P4501A2 are downregulated at both transcriptional and post-transcriptional levels by conditions resulting in interferon-alpha/beta induction.

The interferon mediated downregulation of constitutive and inducible cytochrome P450 enzymes occurs through a pretranslational mechanism which depresses the mRNA encoding cytochrome P450s. We measured the transcription rates of CYP1A genes and the turnover of CYP1A mRNA in rats treated with the interferon-alpha/beta inducer polyinosinic acid-polycytidylic acid. The rate of transcription of CYP1A1 and CYP1A2 genes was significantly decreased in hepatic nuclei isolated from male rats treated with polyinosinic acid-polycytidylic acid (10 mg/kg). In addition the rate of degradation of hepatic CYP1A1 and CYP1A2 mRNA was examined following the inhibition of de novo transcription by actinomycin D (1 mg/kg). Messenger RNA levels were analysed by Northern and slot blotting with a 1.2 kb murine CYP1A1 cDNA probe. Interferon significantly augmented the rate of loss of CYP1A1 and CYP1A2 mRNAs suggesting that post-transcriptional degradation of mRNA contributes to the pre-translational events that cause cytochrome P450 downregulation. These results support the involvement of both transcriptional and post-transcriptional mechanisms in the loss of cytochrome P450s mediated by interferon inducers.

Animals↗
PubMed1991

Photoinduced electron transfer between cytochrome c peroxidase and yeast cytochrome c labeled at Cys 102 with (4-bromomethyl-4'-methylbipyridine)[bis(bipyridine)]ruthenium2+.

The synthesis of (4-bromomethyl-4'-methylbipyridine) [bis(bipyridine)]ruthenium(II) hexafluorophosphate is described. This new reagent was found to selectively label the single sulfhydryl group at Cys-102 on yeast iso-1-cytochrome c to form the (dimethylbipyridine-Cys-102-cytochrome c)[bis(bipyridine)]ruthenium derivative (Ru-102-cyt c). Excitation of Ru-102-cyt c with a short light flash resulted in formation of excited-state Ru(II*), which rapidly transferred an electron to the ferric heme group to form Fe(II). When the cytochrome c peroxidase compound I (CMPI) was present in the solution, electron transfer from photoreduced Fe(II) in Ru-102-cyt c to the radical site in CMPI was observed. At high ionic strength (100 mM sodium phosphate and 25 mM EDTA, pH 7), second-order kinetics were observed with a rate constant of (7.5 +/- 0.7) x 10(7) M-1 s-1. The second-order rate constant for native iso-1-cytochrome c was (6.7 +/- 0.7) x 10(7) M-1 s-1 under these conditions. The second-order rate constant for electron transfer from Ru-102-cyt c to the radical site in CMPI increased as the ionic strength was decreased, reaching a value of (4.8 +/- 0.5) x 10(8) M-1 s-1 in 40 mM EDTA, pH 7. At lower ionic strength, a complex was formed between Ru-102-cyt c and CMPI, and the rate for intracomplex electron transfer to the radical site was found to be greater than 50,000 s-1.(ABSTRACT TRUNCATED AT 250 WORDS)

Cysteine↗
PubMed1994

Roles of the soluble cytochrome c2 and membrane-associated cytochrome cy of Rhodobacter capsulatus in photosynthetic electron transfer.

Genetic evidence indicates that Rhodobacter capsulatus has two different pathways for reduction of the photooxidized reaction center (RC) [Jenney, F. E., & Daldal, F. (1993) EMBO J. 12, 1283-1292]. One pathway is via the water soluble cytochrome (cyt) c2, and the other is via a novel, membrane-associated c-type cytochrome, cyt cy, now believed to be identical to the cyt cx of Jones et al. [Jones, M. R., et al. (1990) Biochim. Biophys. Acta 975, 59-66] and c354 of Zannoni et al. [Zannoni, D., et al. (1992) Arch. Microbiol. 157, 367-374]. Mutants lacking either cyt c2, cyt cy, or the bc1 complex, as well as various combinations, were utilized to probe the functional role of these cytochromes in electron transfer. Data obtained by monitoring flas induced electron transfer kinetics in the RC, cyt c pool, cyt b, and the carotenoid band shift indicate that there are two pathways for electron transfer from the bc1 complex to the RC in R. capsulatus, one via cyt c2 and the other through cty cy. The two pathways show strikingly different kinetics for RC reduction and cyt c oxidation, and both are present in the wild-type strain MT-1131. After genetic inactivation of both cyt c2 and cyt cy there remains no flash oxidizible c-type cytochrome, and inactivation of cyt cy rather than cyt c2 has a more pronounced effect on the extent of the light-induced membrane potential under the conditions tested.(ABSTRACT TRUNCATED AT 250 WORDS)

Cell Membrane↗
PubMed1991

Electron spin echo envelope modulation spectroscopy supports the suggested coordination of two histidine ligands to the Rieske Fe-S centers of the cytochrome b6f complex of spinach and the cytochrome bc1 complexes of Rhodospirillum rubrum, Rhodobacter sphaeroides R-26, and bovine heart mitochondria.

Electron spin echo envelope modulation (ESEEM) experiments performed on the Rieske Fe-S clusters of the cytochrome b6f complex of spinach chloroplasts and of the cytochrome bc1 complexes of Rhodospirillum rubrum, Rhodobacter sphaeroides R-26, and bovine heart mitochondria show modulation components resulting from two distinct classes of 14N ligands. At the g = 1.92 region of the Rieske EPR spectrum of the cytochrome b6f complex, the measured hyperfine couplings for the two classes of coupled nitrogens are A1 = 4.6 MHz and A2 = 3.8 MHz. Similar couplings are observed for the Rieske centers in the three cytochrome bc1 complexes. These ESEEM results indicate a nitrogen coordination environment for these Rieske Fe-S centers that is similar to that of the Fe-S cluster of a bacterial dioxygenase enzyme with two coordinated histidine ligands [Gurbiel, R. J., Batie, C. J., Sivaraja, M., True, A. E., Fee, J. A., Hoffman, B. M., & Ballou, D. P. (1989) Biochemistry 28, 4861-4871]. The Rieske Fe-S cluster lacks modulation components from a weakly coupled peptide nitrogen observed in water-soluble spinach ferredoxin. Treatment with the quinone analogue inhibitor DBMIB causes a shift in the Rieske EPR spectrum to g = 1.95 with no alteration in the magnetic coupling to the two nitrogen atoms. However, the ESEEM pattern of the DBMIB-altered Rieske EPR signal shows evidence of an additional weakly coupled nitrogen similar to that observed in the spinach ferredoxin ESEEM patterns.

Animals↗
PubMed1985

Resonance raman spectra of CN--bound cytochrome oxidase: spectral isolation of cytochromes a2+, a3(2+), and a3(2+)(CN-).

Reduced cyanide-bound cytochrome oxidase in the absence of any oxygen gives a resonance Raman spectrum consistent with that expected for low-spin heme a. Thus, in contrast to prior reports, ligand binding of cytochrome a3 to form a six-coordinate low-spin ferrous heme does not result in any unusual electronic structure, hydrogen bonding, environment, or conformation of the formyl group. It appears unlikely that there are any changes in this group in cytochrome a3 that control the ligand affinity or redox potential in physiological forms of the ferrous enzyme. With the use of our difference spectrometer and by appropriately selecting the laser excitation frequency, we are able to isolate spectrally cytochromes a2+, a3(2+), and a3(2+)(CN-). The addition of a small amount of oxygen to a preparation of the cyanide-bound reduced enzyme results in a complex with the same Raman spectrum as that previously reported to originate from the cyanide-bound reduced complex. Any oxygen present in the sample leads to enzyme turnover resulting in a mixed valence state [a2+a3(3+)(CN-)]. The comparison between the data on the cyanide-bound reduced enzyme and the data on the CO-bound reduced enzyme illustrates that cyanide binding affects only the modes that respond to the spin state of the ferrous iron, while CO binding affects vibrational modes that respond to a pi-electron density change as well.

Animals↗
PubMed1989

Cytochrome c oxidase: evidence for interaction of water molecules with cytochrome a.

The resonance Raman spectra of cytochrome c oxidase in protonated buffer compared to that in deuterated buffer indicate that water molecules are near the heme of cytochrome a. Differences in widths of the heme line at 1610 cm-1, after short exposure to D2O, and, additionally, of the heme line at 1625 cm-1, after long exposure, can be accounted for by changes in resonance vibrational energy transfer between modes of cytochrome a2+ and the bending mode of water molecules in the heme pocket. On the basis of the assignment of these modes, we place one water molecule near the vinyl group and one water molecule near the formyl group of the cytochrome a heme. These water molecules may play several possible functional roles.

Animals↗