Archives of Biochemistry and Biophysics

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The Isolation and Comparison of Multiple Forms of CYP2B from Untreated and Phenobarbital-Treated Rabbit Liver Microsomes

I. Jansson, J.E. Mole, J.B. Schenkman

Archives of Biochemistry and Biophysics > 1995 > 316 > 1 > 275-284

At low levels of phenobarbital induction two forms of isoenzyme 2 (LM2; CYP2B4) were obtained during purification of cytochrome P450 from rabbit liver microsomes. At high levels of induction only one form (LM2A) was present. Although the two purified forms (LM2A and LM2B) were very similar they differed in: (a) peak elution on CM-Sepharose, (b) wavelength maximum of the reduced P450-CO spectrum, and...

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Methionine Biosynthesis in Higher-Plants .1. Purification and Characterization of Cystathionine γ-Synthase from Spinach Chloroplasts

S. Ravanel, M. Droux, R. Douce

Archives of Biochemistry and Biophysics > 1995 > 316 > 1 > 572-584

Cystathionine γ-synthase, the first enzyme specific for the methionine biosynthetic pathway, was purified to apparent homogeneity from spinach leaf chloroplasts. A nonradioactive assay based on O-phthaldialdehyde derivatization of L-cystathionine and fluorescence detection was developed to determine the cystathionine γ-synthase activity. A unique cystathionine γ-synthase activity was located in the...

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Kinetic Studies of Gluconate Pathway Enzymes from Schizosaccharomyces pombe

C.S. Tsai, J.L. Shi, H.G. Ye

Archives of Biochemistry and Biophysics > 1995 > 316 > 1 > 163-168

Glucose dehydrogenase and gluconate kinase which catalyze two-step reactions of the gluconate pathway have been purified from Schizosaccharomyces pombe. Their steady-state kinetic studies were undertaken. The yeast glucose dehydrogenase requires NADP⁺ as an obligatory coenzyme and mediates the oxidation of D-glucose to D-gluconate via an ordered Bi Bi mechanism with NADP⁺ as the leading substrate...

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The Vitamin D3 Hydroxylase-Associated Protein Is a Propionamide-Metabolizing Amidase Enzyme

R.A. Ettinger, H.F. Deluca

Archives of Biochemistry and Biophysics > 1995 > 316 > 1 > 14-19

Previously we isolated a novel protein that coimmunoprecipitates with the 1,25-dihydroxyvitamin D₃-24R-hydroxylase and 25-hydroxyvitamin D₃-1α-hydroxylase. This kidney-specific protein found in the inner membrane of mitochondria is named the vitamin D₃ hydroxylase-associated protein (VDHAP). To determine a putative function for this protein, an extensive computer search of the deduced amino acid sequence...

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The Interaction of Candida boidinii Formate Dehydrogenase with a New Family of Chimeric Biomimetic Dye-Ligands

N.E. Labrou, Y.D. Clonis

Archives of Biochemistry and Biophysics > 1995 > 316 > 1 > 169-178

Seven chimeric biomimetic dye-ligands (BM) are purpose-designed and synthesized by specific structural modification of the parent anthraquinone dichlorotriazine dye Vilmafix blue A-R (VBAR). Each BM dye is composed of two enzyme-recognition moieties. The terminal biomimetic moiety bears a variable carboxylated structure linked to the triazine ring, thus mimicking the substrate of formate dehydrogenase...

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5-Aminolevulinic Acid Induces Iron Release from Ferritin

P.I. Oteiza, C.G. Kleinman, M. Demasi, E.J.H. Bechara

Archives of Biochemistry and Biophysics > 1995 > 316 > 1 > 607-611

5-Aminolevulinic acid (ALA), a heme precursor accumulated in acute intermittent porphyria (AIP) and lead poisoning, undergoes metal-catalyzed oxidation in air-equilibrated solutions buffered at neutral pH, yielding free radicals (O²⁻, HO^. and ALA^.). The capacity of ALA to release iron from horse spleen and rat liver ferritin in vitro and to concomitantly initiate liposome lipid peroxidation was characterized...

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Characterization of a Dinucleotide-Binding Site in Monoamine Oxidase B by Site-Directed Mutagenesis

S.W. Kwan, D.A. Lewis, B.P. Zhou, C.W. Abell

Archives of Biochemistry and Biophysics > 1995 > 316 > 1 > 385-391

Monoamine oxidase B (MAO B), an integral protein of the outer mitochondrial membrane, catalyzes the oxidative deamination of neuroactive and vasoactive amines. The oxidation step is coupled to the reduction of an obligatory FAD cofactor. In this study, we have examined the role of one amino acid (Glu³⁴) in human MAO B that is thought to play a crucial role in binding to the 2′-hydroxy group of ribose...

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Expression of Liver Fatty Acid Binding Protein in L-Cells: Plasma Membrane Response to Ethanol

F. Schroeder, T. Hubbell, S.M. Colles, W.G. Wood

Archives of Biochemistry and Biophysics > 1995 > 316 > 1 > 343-352

Expression of liver fatty acid binding protein (L-FABP) in transfected L-cell fibroblasts modifies plasma membrane structure and function [Incerpi et al., 1992, Arch. Biochem. Biophys. 298, 35-42], The effect of L-FABP expression on ethanol induced fluidization of plasma membranes was examined. Ethanol in vitro selectively fluidized the exofacial leaflet of the plasma membranes from L-cells expressing...

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α1-Adrenergic and m3-Muscarinic Receptor Stimulation of Phosphatidylinositol 4,5-Bisphosphate-Specific Phospholipase C Are Independently Mediated by Gαq/11 in Rat Parotid Gland Membranes

K. Sawaki, B.J. Baum, I.S. Ambudkar

Archives of Biochemistry and Biophysics > 1995 > 316 > 1 > 535-540

Stimulation of m₃-muscarinic cholinergic receptors (m₃AChR) in the rat parotid gland increases the hydrolysis of phosphatidylinositol 4, 5-bisphosphate (PIP₂) via activation of the G_αq/11 family of G-proteins (Sawaki et al. (1993) Arch. Biochem. Biophys., 546-550). Herein we report that α₁-adrenergic receptor (α₁-AR) stimulation of PIP₂ hydrolysis is also mediated via α subunits of the G_q/11 family...

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Protein Expression, Selective Isotopic Labeling, and Analysis of Hyperfine-Shifted NMR Signals of Anabaena 7120 Vegetative [2Fe-2S]Ferredoxin

H. Cheng, W.M. Westler, B. Xia, B.H. Oh, more

Archives of Biochemistry and Biophysics > 1995 > 316 > 1 > 619-634

Two alternative T7 RNA promoter/polymerase systems have been employed for the heterologous expression of a plant-type [2Fe-2S]ferredoxin, Anabaena 7120 vegetative ferredoxin, in Escherichia coli at high levels (∼20 mg/liter of culture). One system was used when ¹⁵N-labeling the ferredoxin uniformly by growing E. coli with ¹⁵NH₄Cl as the nitrogen source; the other was used in conjunction with auxotrophic...

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The Hepatocellular Metabolism of 4-Hydroxynonenal by Alcohol Dehydrogenase, Aldehyde Dehydrogenase, and Glutathione S-Transferase

D.P. Hartley, J.A. Ruth, D.R. Petersen

Archives of Biochemistry and Biophysics > 1995 > 316 > 1 > 197-205

It has previously been reported that isolated rat hepatocytes rapidly and completely metabolize high concentrations of 4-hydroxy-2,3-(E)-nonenal (4-HNE). However, until this report, the degree to which oxidative-reductive and nonoxidative metabolic pathways function in the depletion of 4-HNE by isolated rat hepatocytes has been speculative. The objective of the present study was to quantitate the...

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Divalent Cations and Chelators as Regulators of Brain Fructose-1,6-bisphosphatase

S. Chattorajbhattacharyya, A.L. Majumder

Archives of Biochemistry and Biophysics > 1995 > 316 > 1 > 63-69

Purified fish and rat brain FruP₂ase(s) are stimulated by a number of chelators, viz., histidine, EDTA, citrate, imidazole, and a number of histidine analogues. These also impart 5′-AMP sensitivity to the otherwise insensitive enzyme. Beyond 3 mM concentration, histidine inhibits the enzyme activity, which can be prevented by Mn²⁺. Atomic absorption spectrophotometry showed the presence of 5-6 mol...

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δ-Aminolevulinate Increases Heme Saturation and Yield of Human Cystathionine β-Synthase Expressed in Escherichia coli

V. Kery, D. Elleder, J.P. Kraus

Archives of Biochemistry and Biophysics > 1995 > 316 > 1 > 24-29

We recently expressed human cystathionine β-synthase (CBS) in Escherichia coli and purified it to homogeneity. We showed that CBS requires heme in addition to pyridoxal 5′-phosphate for its function. Previously, CES, only about 20% saturated with heme, was purified from transformed bacteria. In the present study, we supplemented the bacteria with 0.3 mM δ-aminolevulinate (δALA), a precursor of heme...

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Dynamics of Xanthine Oxidase- and Fe3+-ADP-Dependent Lipid Peroxidation in Negatively Charged Phospholipid Vesicles

K. Fukuzawa, K. Soumi, M. Iemura, S. Goto, more

Archives of Biochemistry and Biophysics > 1995 > 316 > 1 > 83-91

Superoxide (O⁻ ₂)-dependent lipid peroxidation on addition of xanthine oxidase (XO) and Fe³⁺-ADP was induced in egg phosphatidylcholine (PC) liposomes containing dicetylphosphate (DCP), which are negatively charged like biological membranes, but not in uncharged egg PC liposomes. Positively charged Fe³⁺ ADP interacted more with negatively charged egg PC-DCP liposomes than with uncharged egg PC liposomes...

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Metabolism of Epoxyeicosatrienoic Acids by Cytosolic Epoxide Hydrolase: Substrate Structural Determinants of Asymmetric Catalysis

D.C. Zeldin, S.Z. Wei, J.R. Falck, B.D. Hammock, more

Archives of Biochemistry and Biophysics > 1995 > 316 > 1 > 443-451

The metabolism of cis-epoxyeicosatrienoic acids (EETs), methyl cis-epoxyeicosatrienoates, and cis-epoxyeicosanoic acids by cytosolic epoxide hydrolase was studied to identify substrate structural features important for stereoselective metabolism and chiral diol formation. 14(R), 15(S)-, 11(S),12(R)-, and 8(S),9(R)-EET, the predominant enantiomers present endogenously in rat organs, were metabolized...

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Role of Heparan Sulfate in the Terminal Differentiation of Growth Plate Chondrocytes

S.K. Chintala, R.R. Miller, C.A. Mcdevitt

Archives of Biochemistry and Biophysics > 1995 > 316 > 1 > 227-234

Recent studies have demonstrated that basic fibroblast growth factor (bFGF) plays a key role in the terminal differentiation of growth plate chondrocytes during endochondral ossification. In this paper we examined the potential role of heparan sulfate in the regulation of the action of basic fibroblast growth factor (bFGF) in the terminal differentiation of rat growth plate chondrocytes. As rat growth...

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Fructose-1,6-bisphosphatase in Stage VI Frog Oocytes: Evidence for an Active Enzyme in Vivo

A. Preller, V. Guixe, E. Kessi, T. Ureta

Archives of Biochemistry and Biophysics > 1995 > 316 > 1 > 555-560

The characterization of fructose-1,6-bisphosphatase in stage VI oocytes from the frog Caudiverbera caudiverbera, as well as the in vivo activity, is reported. The enzyme has a subunit molecular weight of approximately 43,500, has an apparent K_m value of 17 μM for fructose-1,6-bisP, and is inhibited by substrate concentrations beyond 100 μM. AMP and fructose-2,6-bisP are strong inhibitors of oocyte...

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Stereochemistry of Benzylamine Oxidation by Copper Amine Oxidases

G. Alton, T.H. Taher, R.J. Beever, M.M. Palcic

Archives of Biochemistry and Biophysics > 1995 > 316 > 1 > 353-361

Copper amine oxidases (EC 1.4.3.6) exhibit stereochemical heterogeneity in their reaction specificities. Enzymes isolated from different sources have previously been shown to catalyze the deamination of tyramine and dopamine with abstraction of the pro-R hydrogen at C-1, the pro-S hydrogen, and net nonstereospecific proton abstraction. In this study we report on the stereochemical course of proton...

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Regulation of Cytochrome P4501A1 Gene Expression in Vascular Smooth Muscle Cells Through Aryl Hydrocarbon Receptor-Mediated Signal Transduction Requires a Protein Synthesis Inhibitor

X.L. Ou, K.S. Ramos

Archives of Biochemistry and Biophysics > 1995 > 316 > 1 > 116-122

The present studies were conducted to evaluate the pattern of cytochrome P4501A1 (CYP1A1) gene inducibility in vascular (aortic) smooth muscle cells (SMCs) upon exposure to selected aromatic hydrocarbons. Challenge of randomly cycling or synchronized subcultures of adult quail aortic SMCs with 30 μM benzo[a]pyrene (BaP) or 10 nM 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) for 24 h failed to induce...

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Cloning and Expression in Escherichia coli of a Novel Thioesterase from Arabidopsis thaliana Specific for Long-Chain Acyl-Acyl Carrier Proteins

P. Dormann, T.A. Voelker, J.B. Ohlrogge

Archives of Biochemistry and Biophysics > 1995 > 316 > 1 > 612-618

An Arabidopsis thaliana partial cDNA was previously identified with a sequence similar to the lauroyl-acyl carrier protein (ACP) thioesterase from Umbellularia californica (Grellet et al., 1993, Plant Physiol. Biochem. 31, 599-602). Using this DNA fragment, we isolated a 1.8-kb cDNA coding for a 412-amino-acid preprotein. The deduced amino acid sequence is 51% identical to the lauroyl-ACP thioesterase...

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Archives of Biochemistry and Biophysics

The Isolation and Comparison of Multiple Forms of CYP2B from Untreated and Phenobarbital-Treated Rabbit Liver Microsomes

Methionine Biosynthesis in Higher-Plants .1. Purification and Characterization of Cystathionine γ-Synthase from Spinach Chloroplasts

Kinetic Studies of Gluconate Pathway Enzymes from Schizosaccharomyces pombe

The Vitamin D3 Hydroxylase-Associated Protein Is a Propionamide-Metabolizing Amidase Enzyme

The Interaction of Candida boidinii Formate Dehydrogenase with a New Family of Chimeric Biomimetic Dye-Ligands

5-Aminolevulinic Acid Induces Iron Release from Ferritin

Characterization of a Dinucleotide-Binding Site in Monoamine Oxidase B by Site-Directed Mutagenesis

Expression of Liver Fatty Acid Binding Protein in L-Cells: Plasma Membrane Response to Ethanol

α1-Adrenergic and m3-Muscarinic Receptor Stimulation of Phosphatidylinositol 4,5-Bisphosphate-Specific Phospholipase C Are Independently Mediated by Gαq/11 in Rat Parotid Gland Membranes

Protein Expression, Selective Isotopic Labeling, and Analysis of Hyperfine-Shifted NMR Signals of Anabaena 7120 Vegetative [2Fe-2S]Ferredoxin

The Hepatocellular Metabolism of 4-Hydroxynonenal by Alcohol Dehydrogenase, Aldehyde Dehydrogenase, and Glutathione S-Transferase

Divalent Cations and Chelators as Regulators of Brain Fructose-1,6-bisphosphatase

δ-Aminolevulinate Increases Heme Saturation and Yield of Human Cystathionine β-Synthase Expressed in Escherichia coli

Dynamics of Xanthine Oxidase- and Fe3+-ADP-Dependent Lipid Peroxidation in Negatively Charged Phospholipid Vesicles

Metabolism of Epoxyeicosatrienoic Acids by Cytosolic Epoxide Hydrolase: Substrate Structural Determinants of Asymmetric Catalysis

Role of Heparan Sulfate in the Terminal Differentiation of Growth Plate Chondrocytes

Fructose-1,6-bisphosphatase in Stage VI Frog Oocytes: Evidence for an Active Enzyme in Vivo

Stereochemistry of Benzylamine Oxidation by Copper Amine Oxidases

Regulation of Cytochrome P4501A1 Gene Expression in Vascular Smooth Muscle Cells Through Aryl Hydrocarbon Receptor-Mediated Signal Transduction Requires a Protein Synthesis Inhibitor

Cloning and Expression in Escherichia coli of a Novel Thioesterase from Arabidopsis thaliana Specific for Long-Chain Acyl-Acyl Carrier Proteins

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