Subject(s)
Nutritional Sciences/education , Rural Population , Urban Population , Aged , Books , Dietary Services , Female , Food Labeling , Humans , Interviews as Topic , Male , Periodicals as Topic , Sex Factors , Surveys and Questionnaires , Television , TexasABSTRACT
S-Dimethylarsino-CoA was synthesized by acylation of CoA with dimethylchloroarsine. The new analogue of acetyl-CoA was tested as an active-site-directed irreversible inhibitor of phosphotransacetylase (EC 2.3.1.8), carnitine acetyltransferase (EC 2.3.1.7) and citrate synthase (EC 4.1.3.7). Irreversible inhibition was observed only with phosphotransacetylase, which was derivatized via a simple bimolecular process (k2 = 197 +/- 15 min-1 . M-1). Acetyl-CoA provided complete substrate protection against the inactivation, while phosphate (a substrate) and desulfo-CoA (a competitive inhibitor) provided a partial protection. The inactivation was not reversed by dithiothreitol. The new reagent was a linear competitive inhibitor versus acetyl-CoA with both carnitine acetyltransferase (Ki = 41 microM) and citrate synthase (Ki = 20 microM). Chemical studies showed that S-dimethylarsino-CoA reacts with the thiol of N alpha-acetylcysteine but not with the side-chain functional groups of histidine and lysine. The nature of the chemical modification of cysteine was determined by investigating a model system. Thus the chemical reaction between the thioarsenite linkage of S-dimethylarsinobenzylmercaptan and the thiol of cysteine was shown to involve transesterification of the dimethylarsino group.
Subject(s)
Acetyltransferases/antagonists & inhibitors , Coenzyme A/analogs & derivatives , Phosphate Acetyltransferase/antagonists & inhibitors , Animals , Carnitine O-Acetyltransferase/antagonists & inhibitors , Citrate (si)-Synthase/antagonists & inhibitors , Clostridium/enzymology , Coenzyme A/pharmacology , Columbidae , Magnetic Resonance Spectroscopy , Mathematics , SwineSubject(s)
Acetyl Coenzyme A/metabolism , Affinity Labels/chemical synthesis , Coenzyme A/analogs & derivatives , Acetyl-CoA C-Acetyltransferase/antagonists & inhibitors , Affinity Labels/pharmacology , Chromatography, Thin Layer , Coenzyme A/chemical synthesis , Coenzyme A/pharmacology , Indicators and Reagents , Spectrophotometry, UltravioletSubject(s)
Acetyl Coenzyme A/metabolism , Acetyl-CoA C-Acetyltransferase/antagonists & inhibitors , Acetyl-CoA Hydrolase/antagonists & inhibitors , Acetyltransferases/antagonists & inhibitors , Affinity Labels , Citrate (si)-Synthase/antagonists & inhibitors , Coenzyme A/analogs & derivatives , Oxo-Acid-Lyases/antagonists & inhibitors , Thiolester Hydrolases/antagonists & inhibitors , 3-Hydroxyacyl CoA Dehydrogenases/antagonists & inhibitors , Binding, Competitive , Coenzyme A/metabolism , Coenzyme A/pharmacology , KineticsSubject(s)
Embryo Implantation, Delayed , Embryo Implantation , Pregnancy, Animal , Trypsin/metabolism , Uterus/enzymology , Animals , Female , Fluorometry , Gestational Age , Hydrogen-Ion Concentration , Mice , Pregnancy , TemperatureABSTRACT
Reduced nicotinamide adenine dinucleotide phosphate (NADPH)-nitrate reductase from Neurospora crassa was purified and found to be stimulated by certain amino acids, citrate, and ethylenediaminetetraacetic acid (EDTA). Stimulation by citrate and the amino acids was dependent upon the prior removal of EDTA from the enzyme preparations, since low quantities of EDTA resulted in maximal stimulation. Removal of EDTA from enzyme preparations by dialysis against Chelex-containing buffer resulted in a loss of nitrate reductase activity. Addition of alanine, arginine, glycine, glutamine, glutamate, histidine, tryptophan, and citrate restored and stimulated nitrate reductase activity from 29- to 46-fold. The amino acids tested altered the Km of NADPH-nitrate reductase for NADPH but did not significantly change that for nitrate. The Km of nitrate reductase for NADPH increased with increasing concentrations of histidine but decreased with increasing concentrations of glutamine. Amino acid modulation of NADPH-nitrate reductase activity is discussed in relation to the conservation of energy (NADPH) by Neurospora when nitrate is the nitrogen source.
Subject(s)
Neurospora crassa/enzymology , Neurospora/enzymology , Nitrate Reductases/biosynthesis , Amino Acids/metabolism , Citrates/metabolism , Edetic Acid/metabolism , Enzyme Activation , NADP/metabolism , Neurospora crassa/metabolism , Nitrate Reductases/metabolism , Nitrates/metabolismABSTRACT
Bacillus thuringiensis (ATCC 10792) produces a molybdenum reactive compound (given the trivial name chelin) during growth on iron-deficient medium. This compound accumulates in the culture medium in direct relation to the amount of L-arginine added and reaches a maximum concentration 24 to 48 h after the stationary phase of growth. Chelin absorbs light in the ultraviolet region with absorption maxima at 315 and 248 nm and minima at 284 and 240 nm. Chelin reacts with Na2MoO4, but not with Mo2O4(H2O)6-2+, to form a bright yellow molybdo-chelin complex which absorbs light with an absorption maximum at 330 nm, a minimum at 288 nm, and shoulders at 255 and 400 nm. The differential absorption of molybdo-chelin versus chelin at 425 nm can be used to quantify chelin. This differential absorbance is linear with increasing concentrations of Na2MoO4 and was used to calculate the molar extinction coefficient of molybdochelin at 425 nm (epsilon similar to 6,200). Chelin binds MoO4-2 minus to form a complex (molybdochelin) which migrates as a single band and elutes as a single peak, during acrylamide gel electrophoresis and Sephadex G-15 gel filtration. Molecular weight determinations using Sephadex G-15 gel filtration resulted in an estimated molecular weight of 550 for chelin and an estimated molecular weight of 760 for molybdo-chelin. The peptide nature of chelin is indicated by its positive ninhydrin reaction on thin-layer chromatography plates and by the presence of amino acids in acid-hydrolyzed samples. The major amino acid residues detected were threonine, glycine, and alanine.
