PubMed Health⌕ Search

Biomedical subjects

B Axelrod

Publications and source records attributed to B Axelrod.

At least 19 recordsLinked to original sources
PubMed1998

Neural correlates of levels of emotional awareness. Evidence of an interaction between emotion and attention in the anterior cingulate cortex.

Recent functional imaging studies have begun to identify the neural correlates of emotion in healthy volunteers. However, studies to date have not differentially addressed the brain areas associated with the perception, experience, or expression of emotion during emotional arousal. To explore the neural correlates of emotional experience, we used positron emission tomography (PET) and 15-water to measure cerebral blood flow (CBF) in 12 healthy women during film- and recall-induced emotion and correlated CBF changes attributable to emotion with subjects' scores on the Levels of Emotional Awareness Scale (LEAS), a measure of individual differences in the capacity to experience emotion in a differentiated and complex way. A conjunction analysis revealed that the correlations between LEAS and CBF during film- and recall-induced emotion overlapped significantly (z = 3.74, p < 0. 001) in Brodmann's area 24 of the anterior cingulate cortex (ACC). This finding suggests that individual differences in the ability to accurately detect emotional signals interoceptively or exteroceptively may at least in part be a function of the degree to which the ACC participates in the experiential processing and response to emotion cues. To the extent that this finding is consistent with the functions of the ACC involving attention and response selection, it suggests that this neural correlate of conscious emotional experience is not exclusive to emotion.

Adolescent↗
PubMed1996

Crystal structure of soybean lipoxygenase L-1 at 1.4 A resolution.

Lipoxygenases, which are widely distributed among plant and animal species, are Fe-containing dioxygenases that act on lipids containing (Z,Z)-pentadiene moieties in the synthesis of compounds with a variety of functions. Utilizing an improved strategy of data collection, low temperature, and synchrotron radiation of short wavelength, the structure of ferrous soybean lipoxygenase L-1, a single chain protein of 839 amino acid residues, has been determined by X-ray crystallography to a resolution of 1.4 A. The R-factor for the refined model is 19.7%. General features of the protein structure were found to be consistent with the results of prior crystallographic studies at lower (2.6 A) resolution. In contrast to the prior studies, the binding of a water molecule to the active site Fe was established. The octahedral coordination sphere of the Fe also includes the side chains of His499, His504, His690, and Asn694 as well as the terminal carboxylate of Ile839, which binds as a monodentate ligand. Asn694 is involved in a number of labile polar interactions with other protein groups, including an amide-aromatic hydrogen bond, and appears to be a weak ligand. Several possible access routes for dioxygen and fatty acids to the internal active site and substrate binding cavity are described. The protein structure restricts access to the Fe site such that the formation of an organo-Fe intermediate seems improbable. Structural restrictions pertinent to other proposed reaction intermediates, such as planar pentadienyl and nonplanar allyl radicals, are also discussed.

Crystallography, X-Ray↗
PubMed1995

Crystallization and preliminary X-ray investigation of lipoxygenase-3 from soybeans.

Soybean lipoxygenase-3 has been crystallized by the vapor diffusion method in 16-20% polyethylene glycol (average M(r), 3,400), 0.2 M sodium acetate buffer, pH 5.7, at 21 degrees C, at a protein concentration of 8-15 mg/mL. The crystals, which diffract to 3-A spacings, belong to the monoclinic space group C2. Cell constants are a = 111.9, b = 136.4, and c = 61.6 A and beta = 95.7 degrees. The calculated value of Matthews's constant, Vm = 2.48 A3/kDa, is consistent with the presence of one molecule of lipoxygenase per crystallographic asymmetric unit (Z = 4).

Amino Acid Sequence↗
PubMed1994

Effects of non-right-handedness on risk for sudden death associated with coronary artery disease.

The hypothesis that non-right-handedness is associated with sudden cardiac death was tested based on evidence that sympathetic imbalance may contribute to ventricular arrhythmogenesis and evidence that left-handers may have a shorter lifespan than right-handers. The study included 26 patients with coronary artery disease (CAD), a history of ventricular tachycardia-ventricular fibrillation (VT-VF), and implanted defibrillators, and 26 patients with CAD and no history of serious arrhythmias who were matched for age, sex, and New York Heart Association functional class. Patients with any history of neurologic disorders were excluded. Left-handers either wrote with the left hand or were converted from left- to right-handedness in childhood. Non-right-handers used the left hand for writing, drawing, or throwing. Handedness rates in patients with VT-VF and case-control subjects were compared with published norms in the general population to take expected rates into account. The rates of left-handedness (6 of 26 or 23.1%) and non-right-handedness (9 of 26 or 34.6%) in patients with VT-VF were significantly higher (p < 0.003 and p < 0.0001, 2-tailed, respectively) than those of similarly aged adults in the general population (left-handedness, 5%; non-right-handedness, 10.2%). The rates of left-handedness (2 of 26 or 7.7%) and non-right-handedness (4 of 26 or 15.4%) observed in the case-control group correspond closely to the expected values for that group (left-handedness, 1.3 of 26 or 5%; non-right-handedness, 2.65 of 26 or 10.2%) derived from the general population rates and were not significantly different from them.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗
PubMed1993

Crystallographic determination of the active site iron and its ligands in soybean lipoxygenase L-1.

Five ligands of the active site iron atom in soybean lipoxygenase L-1 have been identified from the electron density map of the crystallized enzyme. The position of the iron atom can be readily and independently located from an anomalous difference electron density map. The ligands identified are His-499, His-504, His-690, Asn-694, and Ile-839, the carboxy-terminal residue. Our previous view that these three histidines are essential for activity and binding of iron, based on site-specific mutation studies, is confirmed. A sixth protein ligand is not present, and the sixth coordination site opens into a wide cleft. The structure of the soybean lipoxygenase was solved by multiple anomalous isomorphous replacements.

Amino Acid Sequence↗
PubMed1992

Identification of the iron-binding histidine residues in soybean lipoxygenase L-1.

Lipoxygenases constitute a class of non-heme, non-sulfur iron dioxygenases acting upon lipids possessing a 1,4-cis-cis-pentadiene moiety. The iron is known to be essential for activity. A motif of six histidine residues has been found in all of the thirteen lipoxygenases, from both plant and animal sources, whose sequences have been reported. We had previously obtained mutant proteins in which each of the 6 conserved histidines of soybean lipoxygenase L-1 had been replaced and found that the mutants H499Q, H504Q (or H504S) and H690Q had no detectable enzymatic activity. We have now found that these inactive proteins contain no Fe, although they have the same electrophoretic mobility as wild-type L-1 under both denaturing and non-denaturing conditions and react with anti-L-1 antibodies.

Amino Acid Sequence↗
PubMed1992

Conserved histidine residues in soybean lipoxygenase: functional consequences of their replacement.

Sequences of 13 lipoxygenases from various plant and mammalian species, thus far reported, display a motif of 38 amino acid residues which includes 5 conserved histidines and a 6th histidine about 160 residues downstream. These residues occur at positions 494, 499, 504, 522, 531, and 690 in soybean lipoxygenase isozyme L-1. Since the participation of iron in the lipoxygenase reaction has been established and existing evidence based on Mössbauer and EXAFS spectroscopy suggests that histidines may be involved in iron binding, the effect of the above residues has been examined in soybean lipoxygenase L-1. Six singly mutated lipoxygenases have been produced in which each of the His residues has been replaced with glutamine. Two additional mutants have been constructed wherein the codons for His-494 and His-504 have been replaced by serine codons. All of the mutant lipoxygenases, which were obtained by expression in Escherichia coli, have mobilities identical to that of the wild-type enzyme on denaturing gel electrophoresis and respond to lipoxygenase antibodies. The mutated proteins H499Q, H504Q, H504S, and H690Q are virtually inactive, while H522Q has about 1% of the wild-type activity. H494Q, H494S, and H531Q are about 37%, 8%, and 20% as active as the wild type, respectively. His-517 is conserved in the several lipoxygenase isozymes but not in the animal isozymes. The mutant H517Q has about 33% of the wild-type activity. The inactive mutants, H499Q, H504Q, H504S, and H690Q, become insoluble when heated for 3 min at 65 degrees C, as does H522Q. The other mutants and the wild-type are stable under these conditions.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1990

Crystallization and preliminary X-ray investigation of lipoxygenase 1 from soybeans.

Soybean lipoxygenase 1 has been crystallized by the vapor diffusion method in 8-10% polyethylene glycol (average Mr 3400), 0.2 M sodium acetate buffer, pH 5.2-5.6, at a protein concentration of 6-12 mg/ml. Microseeding was employed to obtain growth of large single crystals. The crystals, which diffract to at least 2.2-A spacings, are monoclinic and of space group P2(1). Cell constants are: a = 95.4, b = 94.2, and c = 50.4 A and beta = 91.4 degrees. The calculated value of Vm (2.41 A3/Da) is consistent with the probable presence of one molecule of lipoxygenase/crystallographic asymmetric unit (Z = 2).

Chemical Phenomena↗
PubMed1988

Primary structure of soybean lipoxygenase L-2.

The nucleotide sequence of soybean lipoxygenase-2 cDNA has been determined, and the complete amino acid sequence of the enzyme has been deduced. Limited direct amino acid sequence data for lipoxygenase-2 protein support this assignment and exclude mRNA representing lipoxygenase-1 and -3. Lipoxygenase 2 has a molecular weight of 97,036 and contains 865 amino acid residues, in contrast to the isozymes, lipoxygenase-1 and -3, which are known to contain 838 and 859 amino acid residues, respectively. Despite significant differences in behavior between these three isozymes, the amino acid sequences of lipoxygenase-1 and -3 are 81 and 74% identical to lipoxygenase-2, respectively. A region of 40 amino acid residues containing a cluster of six histidines and two tyrosines, which is highly conserved in all three isozymes, is discussed as a possible iron-binding region.

Amino Acid Sequence↗
PubMed1987

Primary structure of soybean lipoxygenase-1.

The primary structure of lipoxygenase-1 from soybeans has been determined. The results were deduced from the nucleotide sequence of the complete cDNA for isozyme lipoxygenase-1 and were confirmed by Edman sequencing of selected peptides obtained by CNBr cleavage and supported by carboxyl-terminal sequencing of the intact protein and by partial peptide analysis employing fast atom bombardment-mass spectroscopy. The enzyme has a molecular weight of 94,038 based on its content of 838 amino acids.

Amino Acid Sequence↗
PubMed1986

Singlet oxygen production by soybean lipoxygenase isozymes.

The oxidation of linoleic acid catalyzed by soybean lipoxygenase isozymes was accompanied by 1268 nm chemiluminescence characteristic of singlet oxygen. The recombination of peroxy radicals as first proposed by Russell (Russell, G.A. (1957) J. Am. Chem. Soc. 79, 3871-3877) is a plausible mechanism for the observed singlet oxygen production. Lipoxygenase-3 was the most active isozyme. Under the optimal aerobic conditions of p2H 7, 100 micrograms/ml lipoxygenase-3, 100 microM linoleic acid, 100 microM 13-hydroperoxylinoleic acid, and air-saturated buffer, the yield of singlet oxygen was 12 +/- 0.4 microM or 12% of the amount predicted by the Russell mechanism. High yields of singlet oxygen required the presence of 13-hydroperoxylinoleic acid. Systems containing lipoxygenase-2 and lipoxygenase-3 produced comparable yields of singlet oxygen without added 13-hydroperoxylinoleic acid, since the lipoxygenase-2 served as an in situ source of hydroperoxide. Lipoxygenase-1 was active only at low oxygen concentrations. Its singlet oxygen-producing capacity was greatly increased by the addition of acetone to the system. Lipoxygenase-2 did not produce detectable quantities of singlet oxygen.

Isoenzymes↗
PubMed1985

Inactivation of muscle adenylate kinase by site-specific destruction of tyrosine 95 using potassium ferrate.

Potassium ferrate, an analog of orthophosphate and a potent oxidizing agent, was found to irreversibly inactivate porcine muscle adenylate kinase. Inhibition was prevented by competitive inhibitors or substrates, indicating that the action of ferrate was site-specific. Inactivation was accompanied by the loss of Cys-25 and Tyr-95. P1,P5-di(adenosine 5')-pentaphosphate (10(-7) M), a powerful competitive inhibitor, gave 50% protection to the enzyme from ferrate inactivation. No loss of tyrosine or cysteine residues was observed under conditions of total protection. The degree of inactivation was proportional to the amount of Tyr-95 destroyed. However, Cys-25 was totally oxidized when only 55% inactivation had occurred. Partially inactivated enzyme exhibited a Km for ATP and AMP similar to that of the untreated enzyme. It appears that Cys-25 may be proximate to a phosphate-binding site but is not directly involved in the catalytic reaction. The results suggest that Tyr-95 is located in the vicinity of a phosphate-binding region of adenylate kinase and is essential for enzyme activity.

Adenylate Kinase↗
PubMed1985

Inactivation of the NADH-dependent activities of nitrate reductase by ferrate.

Nitrate reductase (EC 1.6.6.1) from Chlorella vulgaris, a flavin-cytochrome-molybdenum enzyme, catalyses two types of partial reactions: reduction of exogenous cytochrome c by NADH and reduction of nitrate to nitrite by reduced methyl viologen (reduced 1,1'-dimethyl-4,4'-dipyridine dichloride). Ferrate, an analogue of orthophosphate acting on the phosphate-binding region of the enzymes, abolishes the NADH-nitrate reductase as well as the NADH-cytochrome c activities. In addition, the ability of NADH to reduce the endogenous cytochrome b component of the enzyme is also impaired. The reduction of nitrate by reduced methyl viologen is only partially affected. The results indicate that the ferrate primarily disrupts the NADH site.

Bacterial Proteins↗
PubMed1984

Effect of ferrate, a site-specific phosphate analog, on human deoxyhemoglobin.

Ferrate ion, a phosphate analog and a potent oxidizing agent, is known to inactivate a number of enzymes which interact with phosphoryl compounds. In contrast, enzymes which do not interact with phosphoryl compounds are not affected by comparable concentrations of ferrate. To further explore the specificity of ferrate as a reagent which is specific for phosphoryl binding sites, a study of its effect on human hemoglobin A was undertaken. In the deoxy form, this protein is known to interact with 2,3-bisphosphoglycerate, its natural allosteric inhibitor of cooperative binding of oxygen, while as oxyhemoglobin it does not interact with the inhibitor. Treatment with ferrate ion caused the loss of approximately three amino acid residues per beta chain of human deoxyhemoglobin, His-2, His-143, and Tyr-145, and one residue, presumably Tyr-42, per alpha chain. Oxyhemoglobin was not affected by the reagent. 2,3-Bisphosphoglycerate was found to protect deoxyhemoglobin from the action of ferrate. His-2 and His-143 are among the residues reported to be implicated in the binding of 2,3-bisphosphoglycerate by deoxyhemoglobin [A. Arnone (1972) Nature (London) 237, 146-148].

2,3-Diphosphoglycerate↗