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L H Jones

Publications and source records attributed to L H Jones.

At least 37 records · Page 2Linked to original sources
PubMed1995

Roles of dipolar effects and local charge in the ionic strength dependence of redox reactions between c-type cytochromes.

Redox reactions between different c-type cytochromes were monitored by stopped-flow spectroscopy. Second-order rate constants were determined at different ionic strengths for the reactions of Paracoccus denitrificans cytochrome c-551i with its physiologic redox partner cytochrome c-550, and with the nonphysiologic partner horse heart cytochrome c. The latter two cytochromes are structurally quite similar and exhibit identical redox potentials but bear net charges of -7 and +7, respectively. Despite these opposite overall charges, the ionic strength dependencies for the reaction of each with the acidic cytochrome c-551i were very similar. The observed decrease in reaction rate with increasing ionic strength that was observed with cytochromes c-550 and c-551i, the latter of which bears a net charge of -20, cannot be explained simply on the basis of monopole-monopole interactions. These data were analyzed by two different methods: one which treats proteins as both monopoles and dipoles and considers the net charge; and another which neglects dipolar effects and considers only the local charge of the reactive site of the protein rather than net charge. The applicability of each method to the analysis of these data and to protein electrostatic interactions in general is discussed.

Bacterial Proteins↗
PubMed1995

Effects of indole-3-acetic acid on croton oil- and arachidonic acid-induced mouse ear edema.

The indole-3-acetic acid (IAA) is a plant growth hormone (auxin) being considered as a tryptophan metabolite in animals. The main purpose of this work was to verify IAA's topical anti-inflammatory action using croton oil- or arachidonic acid-induced mouse ear edema, in comparison to known anti-inflammatory agents. IAA antioxidant activity was also verified by measuring the inhibition of brain homogenate lipid peroxidation with the thiobarbituric acid reactive substances (TBARS) test. IAA inhibited the action of both croton oil-and arachidonic acid-induced edema in a dose-dependent manner (4.0 mumoles IAA inhibited 75.8% in croton oil and 82.5% in arachidonic acid induced ear edema). Both IAA (5.3 mM) and indomethacin (8.0 mM) inhibited TBARS formation. Data suggest that IAA exhibits antiinflammatory effect possibly by its anti-oxidant activity.

Administration, Topical↗
PubMed1994

Aromatic amine dehydrogenase, a second tryptophan tryptophylquinone enzyme.

Aromatic amine dehydrogenase (AADH) catalyzes the oxidative deamination of aromatic amines including tyramine and dopamine. AADH is structurally similar to methylamine dehydrogenase (MADH) and possesses the same tryptophan tryptophylquinone (TTQ) prosthetic group. AADH exhibits an alpha 2 beta 2 structure with subunit molecular weights of 39,000 and 18,000 and with a quinone covalently attached to each beta subunit. Neither subunit cross-reacted immunologically with antibodies to the corresponding subunits of MADH, and the N-terminal amino acid sequence of the beta subunit of AADH exhibited no homology with the highly conserved beta subunits of MADH. The absorption spectra for the oxidized, semiquinone, and reduced forms of AADH have been characterized, and extinction coefficients for the absorption maxima of each redox form have been determined. These spectra are very similar to those for MADH, indicating the likelihood of a TTQ cofactor. This was verified by the near identity of the vibrational frequencies and intensities in the resonance Raman spectra for the oxidized forms of AADH and MADH. A stable semiquinone of AADH could be observed during a reductive titration with dithionite, whereas titration with tyramine proceeded directly from the oxidized to the reduced form. AADH was very stable against denaturation by heat and exposure to guanidine. The individual subunits could be separated by gel filtration after incubation in guanidine hydrochloride, and partial reconstitution of activity was observed on recombination of the subunits. Steady-state kinetic analysis of AADH yielded a Vmax of 17 mumol/min/mg and a Km for tyramine of 5.4 microM. Substrate inhibition by tyramine was observed. AADH was irreversibly inhibited by hydrazine, phenylhydrazine, hydroxylamine, semicarbazide, and aminoguanidine. Isonicotinic acid hydrazide (isoniazid) and isonicotinic acid 2-isopropyl hydrazide (iproniazid) were reversible noncompetitive inhibitors of AADH and exhibited K(i) values of 8 and 186 microM, respectively. The similarities and differences between AADH and other amine oxidizing enzymes are also discussed.

Alcaligenes↗
PubMed1993

Binding constants for a physiologic electron-transfer protein complex between methylamine dehydrogenase and amicyanin. Effects of ionic strength and bound copper on binding.

Two soluble proteins, methylamine dehydrogenase and amicyanin, form a physiologically relevant complex in which intermolecular electron transfer occurs. To characterize and quantitate the binding of these two weakly-associating proteins, an ultrafiltration binding assay has been developed which involves brief centrifugation of mixtures of proteins in centrifuge concentrators followed by quantitation of proteins on each side of the filtration membrane by HPLC. Under low ionic strength conditions which are optimal for the redox reaction between these proteins, a Kd of 4.5 microM was measured for the methylamine dehydrogenase-amicyanin complex. The Kd increased by 8-fold in the presence of added salt. Apoamicyanin, which is known from crystallographic analysis to be structurally very similar to amicyanin, exhibited a much higher Kd and much less specific binding than did the holoprotein. Apoamicyanin also exhibited apparent self-association at low ionic strength which was not observed with amicyanin. These observations are correlated with the known crystal structures of these proteins, free and in complex, and with the available biochemical information on the interactions of these two proteins.

Apoproteins↗
PubMed1993

Deuterium kinetic isotope effect and stopped-flow kinetic studies of the quinoprotein methylamine dehydrogenase.

Stopped-flow kinetic studies of the reductive half-reaction of methylamine dehydrogenase from Paracoccus denitrificans yielded kinetic constants for the reversible formation of the imine intermediate formed between the substrate and the tryptophan tryptophylquinone (TTQ) prosthetic group and for the hydrogen abstraction step which occurs concomitantly with TTQ reduction. When CD3NH2 was used as a substrate, deuterium kinetic isotope effects of 4.2 and 3.8, respectively, were measured for the rate constants that correspond to the formation and dissociation of the iminoquinone intermediate. A deuterium kinetic isotope effect of 17.2 was measured for the hydrogen abstraction step. The maximum deuterium kinetic isotope effect which was measured in steady-state kinetic experiments was 3.0. These data are discussed in relation to the reaction mechanism of methylamine dehydrogenase and the similar large deuterium kinetic isotope effect for hydrogen abstraction which has been observed for another quinoprotein, plasma amine oxidase.

Deuterium↗
PubMed1992

Factors affecting the stability of methanol dehydrogenase from Paracoccus denitrificans.

Methanol dehydrogenase from Paracoccus denitrificans was purified to homogeneity in two steps from the periplasmic fraction of methanol-grown cells. The enzyme was composed of subunits of M(r) 67,000 and 12,000, and non-covalently bound pyrroloquinoline quinone. It exhibited a pH optimum at pH values of 9.0 and above. It was not stable at pH greater than 9.0, but exhibited little loss of activity after prolonged incubation at pH values as low as 4.5. Methyl dehydrogenase was relatively stable to thermal denaturation. The thermal stability was enhanced by the presence of Ca2+ and diminished by the presence of EDTA. These data suggest a structural role for Ca2+ in this enzyme, similar to what has been observed with quinoprotein glucose and ethanol dehydrogenases.

Alcohol Oxidoreductases↗
PubMed1992

Cofactor-directed inactivation by nucleophilic amines of the quinoprotein methylamine dehydrogenase from Paracoccus denitrificans.

Phenylhydrazine, semicarbazide, aminoguanidine, hydrazine, and hydroxylamine each irreversibly inactivated methylamine dehydrogenase from Paracoccus denitrificans and caused changes in the absorbance spectrum of the protein-bound tryptophan tryptophylquinone [TTQ] prosthetic group. Different spectral perturbations were observed on reaction with each of these inactivators. In each case a stoichiometry of 2 mol per mol of enzyme (1:1 per cofactor) was required to observe complete modification of the absorbance spectrum. Identical changes were observed in the presence and absence of oxygen. The reactions of hydrazine and hydroxylamine were very rapid, with stoichiometric inactivation occurring in less than 30 s. Inactivation by phenylhydrazine and semicarbazide exhibited apparent bimolecular kinetics and second order rate constants for inactivation, respectively, of 25 min-1 mM-1 and 39 min-1 mM-1. In contrast, inactivation by aminoguanidine exhibited saturation behavior and kinetic parameters of KI = 2.5 mM and kinact = 0.5 min-1 were obtained. Ammonium salts did not inactivate the enzyme, but were reversible competitive inhibitors with respect to methylamine. A Ki of 20 mM was obtained for ammonium chloride. A mechanism for the reactions of these compounds with the TTQ cofactor of methylamine dehydrogenase is proposed, and the relationship of these data to the mechanisms of interaction of these compounds with o-quinones and other quinoproteins which possess TTQ and other quinone cofactors is discussed.

Amines↗
PubMed1992

Reactions of benzylamines with methylamine dehydrogenase. Evidence for a carbanionic reaction intermediate and reaction mechanism similar to eukaryotic quinoproteins.

It had been previously reported that aromatic amines were not substrates for the bacterial quinoprotein methylamine dehydrogenase. In this study, benzylamine-dependent activity was also not observed in the steady-state assay of this enzyme with the artificial electron acceptor phenazine ethosulfate (PES). Benzylamines did, however, stoichiometrically reduce the protein-bound tryptophan tryptophylquinone (TTQ) prosthetic group and acted as reversible competitive inhibitors of methylamine oxidation when the enzyme was assayed with PES. When methylamine dehydrogenase activity was monitored using a steady-state assay which employed its physiological electron acceptor amicyanin instead of PES, very low but detectable benzylamine-dependent activity was observed. The reactions of a series of para-substituted benzylamines with methylamine dehydrogenase were examined. A Hammett plot of the log of Ki values for the competitive inhibition by these amines against sigma p exhibited a negative slope. Rapid kinetic measurements allowed the determination of values of k3 and Ks for the reduction of TTQ by each of these amines. A Hammett plot of log k3 versus sigma p exhibited a positive slope, which suggests that the oxidation of these amines by methylamine dehydrogenase proceeds through a carbanionic reaction intermediate. A negative slope was observed for the correlation between log Ks and sigma p. Plots of log k3 and log Ks against substituent constants which reflected either resonance or field/inductive parameters for each para substituent indicated that the magnitude of k3 was primarily influenced by field/inductive effects while Ks was primarily influenced by resonance effects. No correlation was observed between either k3 or Ks and the relative hydrophobicity of the para-substituted benzylamines or steric parameters.(ABSTRACT TRUNCATED AT 250 WORDS)

Anions↗
PubMed1991

Inhibition by cyclopropylamine of the quinoprotein methylamine dehydrogenase is mechanism-based and causes covalent cross-linking of alpha and beta subunits.

Cyclopropylamine acted as a mechanism-based inhibitor of the quinoprotein methylamine dehydrogenase from Paracoccus denitrificans. The protein-bound quinone cofactor of this enzyme was rapidly reduced by addition of a stoichiometric amount of cyclopropylamine, but this compound did not serve as a substrate for the enzyme in the steady-state kinetic assay. Time-dependent inactivation of the enzyme by cyclopropylamine was observed only in the presence of a reoxidant. Saturation behavior was observed, and values of KI of 3.9 microM and K(inact) of 1.7 min-1 were determined. Enzyme inactivation was irreversible, as no restoration of activity was evident after gel filtration of methylamine dehydrogenase which had been incubated with cyclopropylamine in the presence of a reoxidant. The inactivated enzyme exhibited an altered absorption spectrum. Electrophoretic analysis of inactivated methylamine dehydrogenase indicated that covalent cross-linking of the alpha and beta subunits of this alpha 2 beta 2 oligomeric enzyme had occurred and that the quinone cofactor had been modified. A mechanism for this inhibition is proposed which is based upon the data presented and is consistent with the available structural information on methylamine dehydrogenase.

Binding Sites↗
PubMed1990

pH-dependent semiquinone formation by methylamine dehydrogenase from Paracoccus denitrificans. Evidence for intermolecular electron transfer between quinone cofactors.

The quinonoid confactors of Paracoccus denitrificans methylamine dehydrogenase exhibited a pH-dependent redistribution of electrons from the 50% reduced plus 50% oxidized to the 100% semiquinone redox form. This phenomenon was only observed at pH values greater than 7.5. The semiquinone was not readily reduced by addition of methylamine, consistent with the view that this substrate donates two electrons at a time to each cofactor during catalysis. Once formed at pH 9.0, no change in redox state from 100% semiquinone was observed when the pH was shifted to 7.5, suggesting that the requirement of high pH was for formation and not stability of the semiquinone. The rate of semiquinone formation exhibited a first-order dependence on the concentration of methylamine dehydrogenase, indicating that this phenomenon was a bimolecular process involving intermolecular electron transfer between reduced and oxidized cofactors. The rate of semiquinone formation decreased with decreasing ionic strength, suggesting a role for hydrophobic interactions in facilitating electron transfer between methylamine dehydrogenase molecules. Methylamine dehydrogenase was covalently modified with norleucine methyl ester in the presence of 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide (EDC). This modification did not affect the catalytic activity of the enzyme but greatly inhibited the intermolecular redistribution of electrons at high pH. This modification also prevented subsequent cross-linking by EDC of the large subunit of methylamine dehydrogenase to amicyanin, the natural electron acceptor for this enzyme.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacterial Proteins↗
PubMed1989

Continuity of care. Development and implementation of a shared patient data base.

Although the inpatient Oncology Unit, the Medical Oncology Clinic, and Radiation Oncology provided care for many of the same patients, there was no mechanism for sharing nursing information, and little colleague input from one area to another. In order to meet this need, a nurse from each of the clinic areas was added to the inpatient unit's Patient Care Evaluation Committee. Working through this committee, these nurses developed an Inpatient/Outpatient Data Flow Sheet, which could be initiated in any oncology area to implement information flow when a patient was to be seen in a different setting. It proved to be an effective tool. The flow sheet, along with our rationale, was then presented for consideration as a computerized program to be used between the three areas. After careful investigation, it was approved. This provided the oncology areas with the first data storage capability for nursing in the hospital. It offered oncology nurses in distinct and separate areas access to obtain and update information on shared patients. This manuscript will focus on the computer program and the data base designed for the oncology department and its impact on nurses and patients.

Cancer Care Facilities↗
PubMed1988

HIV infection: educational programs and policies for school personnel.

Acquired Immunodeficiency Syndrome (AIDS) challenges schools as the frontline defense against this epidemic. School personnel should become knowledgeable about human immunodeficiency virus (HIV) infection and get involved in presenting quality HIV/AIDS education programs. This paper provides an epidemiological update on AIDS as it relates to the school-age population. Evidence establishing modes of transmission is emphasized, and recommendations are presented for handling students infected with HIV. Information is provided to assist school staff implement HIV/AIDS education programs, including core concepts to be emphasized.

Acquired Immunodeficiency Syndrome↗
PubMed1983

Plant cell cloning and culture products.

Biotechnological developments in the use of plant cells as sources of biochemicals have now brought several applications within commercial range. Plant propagation has developed faster commercially, but now requires similar biotechnology to enable the automated production of large numbers of plants at low cost. Although some of the enabling technology has been developed for pregerminated seeds this has not been coupled to production of plantlets in bioreactors. Our present lack of knowledge of how to control gene expression in plant cells, which stems from our ignorance of the organization and regulation of the plant genome, is a critical factor limiting all applications of plant cell cultures.

Agriculture↗
PubMed1981

Transfer ribonucleic Acid modification and its relationship to tumorous and nontumorous plant growth.

Detailed analyses of tRNA hydrolysates from four tissue types of Nicotiana tabacum, pith from intact plants, pith growing in culture, habituated tissue in culture, and crown gall tumor tissue in culture, revealed significant qualitative and quantitative differences in the pattern of methylation. Although pith from intact plants and pith growing in culture possessed seven different methylated nucleosides, only two were found in habituated and tumorous tissues in culture. Four of the five compounds accounting for the difference were tentatively identified as methylated guanosines. Evaluation of results in terms of several parameters, including growth rate, the tumorous state, habituation, tissue culture, and potential for differentiation, indicate that the extent of tRNA methylation may be correlated with the potential for differentiation of a particular tissue.

Journal Article↗
PubMed1981

Hemodynamics of the femoral head.

UNLABELLED: We have determined the pressure-flow relationships of the canine femoral head during venous tamponade of the hip capsule. Intra-osseous pressures were determined before and after infusion of the joint with saline solution to a pressure of sixty-five centimeters of water. Femoral head blood flow was simultaneously determined by the indicator-dilution technique utilizing isotopically labeled microspheres. In puppies, pressure in the femoral head rose 248 per cent while flow dropped by 60 per cent after inflation of the capsule. In adults, no statistically significant change in either pressure or flow was seen. Thus, in the immature animal, venous tamponade results in increased pressure and decreased blood flow. In the mature animal, venous tamponade does not alter intra-osseous pressures or blood-flow rates due to the intact intramedullary venous drainage of the adult femoral head. CLINICAL RELEVANCE: In the immature individual, venous tamponade may well be involved in the development of Legg-Perthes disease. A bout of nonspecific synovitis may elevate intracapsular pressure sufficiently to obstruct venous outflow. This creates an increase in intra-osseous pressure and a decrease in femoral head blood flow. Hemodynamic changes of this magnitude have not been shown to induce Legg-Perthes disease; however, a strong suspicion exists that such alterations may be linked to the disease. In the adult, venous tamponade probably is not involved in the pathogenesis of avascular necrosis. The maintenance of intramedullary venous drainage of the epiphysis into the metaphysis may account for the fact that vascular necrosis of the femoral head rarely develops in adult patients with osteoarthritis or rheumatoid arthritis plus an inflammatory synovitis of the hip.

Animals↗