Cytochrome P450 IIIA1 (P450p) requires cytochrome b5 and phospholipid with unsaturated fatty acids.

Abstract:

:In contrast to other P450 enzymes purified from rat liver microsomes, purified P450 IIIA1 (P450p) is catalytically inactive when reconstituted with NADPH-cytochrome P450 reductase and the synthetic lipid, dilauroylphosphatidylcholine. However, purified P450 IIIA1 catalyzes the oxidation of testosterone when reconstituted with NADPH-cytochrome P450 reductase, cytochrome b5, an extract of microsomal lipid, and detergent (Emulgen 911). The present study demonstrates that the microsomal lipid extract can be replaced with one of several naturally occurring phospholipids, but not with cholesterol, sphingosine, sphingomyelin, ceramide, cerebroside, or cardiolipin. The ratio of the testosterone metabolites formed by purified P450 IIIA1 (i.e., 2 beta-, 6 beta-, and 15 beta-hydroxytestosterone) was influenced by the type of phospholipid added to the reconstitution system. The ability to replace microsomal lipid extract with several different phospholipids suggests that the nature of the polar group (i.e., choline, serine, ethanolamine, or inositol) is not critical for P450 IIIA1 activity, which implies that P450 IIIA1 activity is highly dependent on the fatty acid component of these lipids. To test this possibility, P450 IIIA1 was reconstituted with a series of synthetic phosphatidylcholines. Those phosphatidylcholines containing saturated fatty acids were unable to support testosterone oxidation by purified P450 IIIA1, regardless of the acyl chain length (C6 to C18). In contrast, several unsaturated phosphatidylcholines supported testosterone oxidation by purified P450 IIIA1, and in this regard dioleoylphosphatidylcholine (PC(18:1)2) was as effective as microsomal lipid extract and naturally occurring phosphatidylcholine or phosphatidylserine. These results confirmed that P450 IIIA1 activity is highly dependent on the fatty acid component of phospholipids. A second series of experiments was undertaken to determine whether microsomal P450 IIIA1, like the purified enzyme, is dependent on cytochrome b5. A polyclonal antibody against purified cytochrome b5 was raised in rabbits and was purified by affinity chromatography. Anti-cytochrome b5 caused a approximately 60% inhibition of testosterone 2 beta-, 6 beta-, and 15 beta-hydroxylation by purified P450 IIIA1 and inhibited these same reactions by approximately 70% when added to liver microsomes from dexamethasone-induced female rats. Overall, these results suggest that testosterone oxidation by microsomal cytochrome P450 IIIA1 requires cytochrome b5 and phospholipid containing unsaturated fatty acids.

journal_name

Arch Biochem Biophys

authors

Eberhart DC,Parkinson A

doi

10.1016/0003-9861(91)90128-6

subject

Has Abstract

pub_date

1991-12-01 00:00:00

pages

231-40

issue

2

eissn

0003-9861

issn

1096-0384

pii

0003-9861(91)90128-6

journal_volume

291

pub_type

杂志文章
  • Regulation of glycan processing by Golgi enzymes from red kidney bean (Phaseolus vulgaris) seedlings.

    abstract::The effect of a protein matrix on the processing of glycoprotein glycans by Golgi enzymes from plant seedlings has been determined with an artificial glycoprotein system, comparing the processing rates of glycan-(biotinyl)Asn (or glycan-(biotinamidohexanoyl)Asn) substrates either free or bound to avidin. An analysis o...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1016/0003-9861(91)90020-j

    authors: Lu Y,Wold F

    更新日期:1991-04-01 00:00:00

  • Raman spectroscopic evidence for a disulfide bridge in calf gamma II crystallin.

    abstract::Laser Raman spectroscopy has been applied to native and dithiothreitol-treated bovine cortical gamma II crystallin to examine the state of the thiol groups and the presence of a putative disulfide bridge. The data reveal significant differences in two key spectral regions. In the thiol stretching region (2500-2600 cm-...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1016/0003-9861(89)90106-9

    authors: Pande J,McDermott MJ,Callender RH,Spector A

    更新日期:1989-02-15 00:00:00

  • Binding properties of glycosaminoglycans to lysozyme--effect of salt and molecular weight.

    abstract::The cationic protein, lysozyme, is present in cartilage but its precise role in this tissue has not yet been established. This study shows that two major and structurally similar glycosaminoglycans (GAGs) of cartilage, i.e., chondroitin sulfate (CS) and hyaluronan (HA) interact with lysozyme at salt concentrations up ...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1006/abbi.1994.1134

    authors: Van Damme MP,Moss JM,Murphy WH,Preston BN

    更新日期:1994-04-01 00:00:00

  • The size and heterogeneity of the messenger RNA associated with 70 S monosomes from down-shifted Escherichia coli.

    abstract::After an energy source shift-down, Escherichia coli accumulates 70 S ribosome-mRNA complexes ("70 S monosomes"). The monosome mRNA strands are predominantly primary transcription products with purine nucleoside 5'-triphosphate and 5'-diphosphate termini present at a 1:2 ratio. The number-average chain length is 564 +/...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1016/0003-9861(85)90367-4

    authors: Wnek AP,Jacobson LA

    更新日期:1985-08-15 00:00:00

  • Vitamin D improves pulmonary function in a rat model for congenital diaphragmatic hernia.

    abstract::A congenital diaphragmatic hernia (CDH) is an anomaly caused by defects in the diaphragm; the resulting limited thorax cavity in turn restricts lung growth (pulmonary hypoplasia). This condition is related to pulmonary hypertension. Despite advances in neonatal CDH therapy, the mortality for severe pulmonary hypoplasi...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1016/j.abb.2021.108769

    authors: Ito Y,Tsuda H,Imai K,Miki R,Miura M,Tachi A,Tano S,Hirako-Takamura S,Moriyama Y,Ushida T,Kobayashi T,Sumigama S,Kajiyama H,Kikkawa F,Kotani T

    更新日期:2021-01-20 00:00:00

  • An arginine residue involved in allosteric regulation of cystathionine β-synthase (CBS) domain-containing pyrophosphatase.

    abstract::Inorganic pyrophosphatase containing a pair of regulatory CBS domains (CBS-PPase1) is allosterically inhibited by AMP and ADP and activated by ATP and diadenosine polyphosphates. Mononucleotide binding to CBS domains and substrate binding to catalytic domains are characterized by positive co-operativity. Bioinformatic...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1016/j.abb.2018.11.024

    authors: Anashkin VA,Orlov VN,Lahti R,Baykov AA

    更新日期:2019-02-15 00:00:00

  • Where is the vitamin D receptor?

    abstract::The vitamin D receptor (VDR) is a member of the nuclear receptor superfamily and plays a central role in the biological actions of vitamin D. VDR regulates the expression of numerous genes involved in calcium/phosphate homeostasis, cellular proliferation and differentiation, and immune response, largely in a ligand-de...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章,评审

    doi:10.1016/j.abb.2012.04.001

    authors: Wang Y,Zhu J,DeLuca HF

    更新日期:2012-07-01 00:00:00

  • Biochemical and immunological identity of the hepatic peroxisomal and microsomal trans-2-enoyl CoA hydratase bifunctional protein.

    abstract::In the present study, the hepatic microsomal and peroxisomal bifunctional trans-2-enoyl CoA hydratases were isolated and purified from rats treated with 2% di-(2-ethylhexyl)phthalate for 8 days. These two enzymes (microsomal and peroxisomal) were purified with the identical purification procedures and had identical mo...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1016/0003-9861(87)90042-7

    authors: Cook L,Ghesquier D,Nagi MN,Favreau LV,Cinti DL

    更新日期:1987-02-01 00:00:00

  • Pancreatic exocrine secretion is blocked by inhibitors of methylation.

    abstract::A number of early experiments suggested a relationship between methyl group metabolism and the exocrine secretion of the pancreas. These included nutritional studies showing that ethionine, the ethyl analog of methionine which inhibits cellular methylation reactions, is a specific pancreatic toxin. Other studies indic...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1006/abbi.1997.0249

    authors: Capdevila A,Decha-Umphai W,Song KH,Borchardt RT,Wagner C

    更新日期:1997-09-01 00:00:00

  • The mouse plasma membrane Ca2+ pump isoform 1 promoter: cloning and characterization.

    abstract::The expression of plasma membrane Ca2+ pump (PMCA) is regulated by various hormones or agonists via multiple second messenger pathways. Two different 5' segments of the PMCA1 gene (isoform 1) were cloned from a mouse genomic library. While one segment contained the 3' end of intron 1 and exon 2, the other segment was ...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1006/abbi.1995.1041

    authors: Du Y,Carlock L,Kuo TH

    更新日期:1995-01-10 00:00:00

  • Inhibition of platelet aggregation by the recombinant cysteine-rich domain of the hemorrhagic snake venom metalloproteinase, atrolysin A.

    abstract::The P-III class of venom metalloproteinases has, in addition to the proteinase domain, a disintegrin-like domain and a cysteine-rich domain. Recent evidence has shown that the nonproteinase domains of the P-III class of hemorrhagic metalloproteinases function in the inhibition of platelet aggregation by blocking essen...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1006/abbi.1999.1517

    authors: Jia LG,Wang XM,Shannon JD,Bjarnason JB,Fox JW

    更新日期:2000-01-01 00:00:00

  • Reactive oxygen species produced by liver mitochondria of rats in sepsis.

    abstract::Reactive oxygen species (ROS) can be generated in experimental shock states through several different mechanisms. We measured ROS production in metabolically active liver mitochondria from rats rendered septic by cecal ligation and puncture. By polarography, the State 4 and State 3 respiration rates of liver mitochond...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1006/abbi.1995.1011

    authors: Taylor DE,Ghio AJ,Piantadosi CA

    更新日期:1995-01-10 00:00:00

  • Low temperature plasma induces angiogenic growth factor via up-regulating hypoxia-inducible factor 1α in human dermal fibroblasts.

    abstract::Numerous studies on the application of low temperature plasma (LTP) have produced impressive results, including antimicrobial, antitumor, and wound healing effects. Although LTP research has branched out to include medical applications, the detailed effects and working mechanisms of LTP on wound healing have not been ...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1016/j.abb.2017.07.012

    authors: Cui HS,Joo SY,Lee DH,Yu JH,Jeong JH,Kim JB,Seo CH

    更新日期:2017-09-15 00:00:00

  • Characterization of the subunits of beta-conglycinin.

    abstract::Four subunits of beta-conglycinin were purified from soybean cultivar CX 635-1-1-1, and were designated alpha, alpha', beta, and beta' in accordance with nomenclature proposed by Thanh and Shibasaki [(1977) Biochim. Biophys. Acta 490, 370-384]. Of these subunits, beta' has not previously been reported or characterized...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1016/0003-9861(85)90787-8

    authors: Coates JB,Medeiros JS,Thanh VH,Nielsen NC

    更新日期:1985-11-15 00:00:00

  • Stimulation of the DNA unwinding activity of human DNA helicase II/Ku by phosphorylation.

    abstract::The Ku autoantigen is a heterodimeric protein of 70- and 83-kDa subunits, endowed with duplex DNA end-binding capacity and DNA helicase activity (Human DNA Helicase II, HDH II). HDH II/Ku is well established as the DNA binding component, the regulatory subunit as well as a substrate for the DNA-dependent protein kinas...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1016/j.abb.2007.11.005

    authors: Ochem AE,Rechreche H,Skopac D,Falaschi A

    更新日期:2008-02-01 00:00:00

  • Regulation and intracellular localization of the biotin holocarboxylase synthetase of 3T3-L1 cells.

    abstract::A quantitative assay has been developed to measure holocarboxylase synthetase activity in cellular extracts. This assay was based on measuring the incorporation of [3H]biotin of high specific activity (4.3 Ci/mmol) into purified rat liver apopyruvate carboxylase. With this assay, holocarboxylase synthetase in 3T3-L1 m...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1016/0003-9861(83)90026-7

    authors: Chang HI,Cohen ND

    更新日期:1983-08-01 00:00:00

  • Redundant enhancement of mouse constitutive androstane receptor transactivation by p160 coactivator family members.

    abstract::Constitutive androstane receptor (CAR) transactivation is enhanced by p160 coactivators, which include three members, SRC-1, SRC-2, and SRC-3. Each of the p160 coactivators enhanced mouse CAR (mCAR) transactivation of the CYP2B1 phenobarbital (PB)-responsive enhancer in transfected cultured cells and mouse hepatocytes...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1016/j.abb.2007.09.005

    authors: Xia J,Liao L,Sarkar J,Matsumoto K,Reddy JK,Xu J,Kemper B

    更新日期:2007-12-01 00:00:00

  • Characterization of rabbit UDP-glucuronosyltransferase UGT1A7: tertiary amine glucuronidation is catalyzed by UGT1A7 and UGT1A4.

    abstract::A rabbit liver UDP-glucuronosyltransferase cDNA that is related to human and rat UGT1A7 has been identified. The predicted amino acid sequence of the UGT1A7l displays 80% similarity to that encoded by human HP4 (UGT1A9), but 81% to that predicted for human UGT1A7 and 77% to the rat UGT1A7 (UGTA2). The exons encoding h...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1006/abbi.1997.0214

    authors: Bruck M,Li Q,Lamb JG,Tukey RH

    更新日期:1997-08-15 00:00:00

  • Evidence for a novel set of small heat-shock proteins that associates with the mitochondria of murine PC12 cells and protects NADH:ubiquinone oxidoreductase from heat and oxidative stress.

    abstract::Several previously unreported small heat-shock proteins (sHsps) were detected in mitochondria from heat-stressed rat PC12 cells, but not in unstressed controls. Functional inactivation of the mitochondrial sHsps with murine Hsp25 antibody indicated that these sHsps protect NADH:ubiquinone oxidoreductase and NADH dehyd...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1006/abbi.1999.1177

    authors: Downs CA,Jones LR,Heckathorn SA

    更新日期:1999-05-15 00:00:00

  • Purification and characterization of glucosamine-6-phosphate deaminase from dog kidney cortex.

    abstract::Glucosamine-6-phosphate deaminase (EC 5.3.1.10) from dog kidney cortex was purified to homogeneity, as judged by several criteria of purity. The purification procedure was based on two biospecific affinity chromatography steps, one of them using N-epsilon-amino-n-hexanoyl-D-glucosamine-6-phosphate agarose, an immobili...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1016/0003-9861(92)90664-i

    authors: Lara-Lemus R,Libreros-Minotta CA,Altamirano MM,Calcagno ML

    更新日期:1992-09-01 00:00:00

  • Matrix metalloproteinase-9 in a unique proteoglycan form in avian embryonic growth plate cartilage.

    abstract::The appearance of a high molecular weight gelatinolytic enzyme (230 kDa) correlated with cartilage collagen loss in chick embryonic tibias cultured with lipopolysaccharide. This 230 kDa enzyme was purified and its activity was measured on synthetic and natural substrates. The enzyme was activated by aminophenylmercuri...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1016/j.abb.2012.02.003

    authors: Patchigolla RK,Knudson W,Schmid TM

    更新日期:2012-04-01 00:00:00

  • Mechanistic analysis of Mycobacterium tuberculosis Rv1347c, a lysine Nepsilon-acyltransferase involved in mycobactin biosynthesis.

    abstract::Mycobactin acylation plays a crucial role in the ability of Mycobacterium tuberculosis to acquire intracellular iron during infection. M. tuberculosis Rv1347c, the lysine N(epsilon)-acyltransferase responsible for mycobactin acylation, represents a valid target for the development of novel anti-tubercular agents. Here...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1016/j.abb.2008.05.013

    authors: Frankel BA,Blanchard JS

    更新日期:2008-09-15 00:00:00

  • Expression and processing of a bacterial endoglucanase in transgenic mice.

    abstract::The C6.5 endoglucanase from Bacillus subtilis catalyzes the hydrolyses of beta-glucans. This enzyme, which is also produced by many ruminant microbes, is not part of the normal digestive repertoire of monogastric animals. We have generated transgenic mice which express the C6.5 endoglucanase gene specifically in the p...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1006/abbi.1999.1243

    authors: Zhang JX,Meidinger R,Forsberg CW,Krell PJ,Phillips JP

    更新日期:1999-07-15 00:00:00

  • Lactococcus lactis TrxD represents a subgroup of thioredoxins prevalent in Gram-positive bacteria containing WCXDC active site motifs.

    abstract::Three protein disulfide reductases of the thioredoxin superfamily from the industrially important Gram-positive Lactococcus lactis (LlTrxA, LlTrxD and LlNrdH) are compared to the "classical" thioredoxin from Escherichia coli (EcTrx1). LlTrxA resembles EcTrx1 with a WCGPC active site motif and other key residues conser...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1016/j.abb.2014.09.010

    authors: Björnberg O,Efler P,Ebong ED,Svensson B,Hägglund P

    更新日期:2014-12-15 00:00:00

  • Purification and characterization of an abscisic acid-inducible anionic peroxidase associated with suberization in potato (Solanum tuberosum).

    abstract::An anionic peroxidase (EC 1.11.1.7), thought to be involved in suberization, was purified 110-fold from wound-healing slices of Solanum tuberosum by a combination of ammonium sulfate fractionation, Sephadex G-100 gel filtration, isoelectric focusing, and phenyl-Sepharose CL-4B chromatography in 24% yield. The purified...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1016/0003-9861(85)90060-8

    authors: Espelie KE,Kolattukudy PE

    更新日期:1985-08-01 00:00:00

  • Reoxygenation injury and antioxidant protection: a tale of two paradoxes.

    abstract::Under certain circumstances, added antioxidants can protect tissues against reoxygenation injury after ischemia. Yet reperfusing blood carries many antioxidants with it. The implications of this "antioxidant paradox" are discussed. ...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章,评审

    doi:10.1016/0003-9861(90)90635-c

    authors: Gutteridge JM,Halliwell B

    更新日期:1990-12-01 00:00:00

  • Effect of glutathione on aconitase in Escherichia coli.

    abstract::The effect of glutathione (GSH) on the superoxide-sensitive [4Fe-4S]-containing aconitase of Escherichia coli was explored. A mutant deficient in GSH biosynthesis, designated gshA, grew slower in a defined medium than did the parental strain and this effect was more pronounced when succinate was supplied as the carbon...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1006/abbi.1993.1120

    authors: Gardner PR,Fridovich I

    更新日期:1993-02-15 00:00:00

  • Phosphatidylinositol modulates the response of calmodulin-dependent phosphatase to calmodulin.

    abstract::Phosphatidylinositol (PtdIns) and many other phospholipids activated calmodulin (CaM)-dependent phosphatase in the presence or absence of Ca2+, and the stimulation was more pronounced in the presence of Ca2+. In addition, PtdIns modulated the response of phosphatase to CaM: at low and nonstimulatory concentrations (le...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1016/0003-9861(89)90005-2

    authors: Huang SL,Merat D,Cheung WY

    更新日期:1989-04-01 00:00:00

  • Conserved active site aspartates and domain-domain interactions in regulatory properties of the sugar kinase superfamily.

    abstract::The structures of the sugar kinase/heat shock 70/actin superfamily of enzymes show that the active site is located in a deep cleft between two domains whose relative movement defines a domain closure conformational change thought to be involved in the catalytic and regulatory properties of members of the superfamily. ...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1006/abbi.1997.0444

    authors: Pettigrew DW,Smith GB,Thomas KP,Dodds DC

    更新日期:1998-01-15 00:00:00

  • Relationship of chloroquine-induced redistribution of a neutral aminopeptidase to hemoglobin accumulation in malaria parasites.

    abstract::To study the relationship between neutral aminopeptidase activity and hemoglobin accumulation in malaria parasites, we treated mice infected with Plasmodium berghei NYU-2 with chloroquine intraperitoneally in doses ranging from 0.3 to 3 micromol per 25 g mouse. Preparations of infected erythrocytes (normalized to repr...

    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章

    doi:10.1016/s0003-9861(02)00688-4

    authors: Fitch CD,Cai GZ,Chen YF,Ryerse JS

    更新日期:2003-02-15 00:00:00