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D C White

Publications and source records attributed to D C White.

At least 19 recordsLinked to original sources
PubMed1995

Movement and force produced by a single myosin head.

Muscle contraction is driven by the cyclical interaction of myosin with actin, coupled to the breakdown of ATP. Studies of the interaction of filamentous myosin and of a double-headed proteolytic fragment, heavy meromyosin (HMM), with actin have demonstrated discrete mechanical events, arising from stochastic interaction of single myosin molecules with actin. Here we show, using an optical-tweezers transducer, that a single myosin subfragment-1 (S1), which is a single myosin head, can act as an independent generator of force and movement. Our analysis accounts for the broad distribution of displacement amplitudes observed, and indicates that the underlying movement (working stroke) produced by a single acto-S1 interaction is approximately 4 nm, considerably shorter than previous estimates but consistent with structural data. We measure the average force generated by S1 or HMM to be at least 1.7 pN under isometric conditions.

Actins↗
PubMed1995

Aromatic-degrading Sphingomonas isolates from the deep subsurface.

An obligately aerobic chemoheterotrophic bacterium (strain F199) previously isolated from Southeast Coastal Plain subsurface sediments and shown to degrade toluene, naphthalene, and other aromatic compounds (J. K. Fredrickson, F. J. Brockman, D. J. Workman, S. W. Li, and T. O. Stevens, Appl. Environ. Microbiol. 57:796-803, 1991) was characterized by analysis of its 16S rRNA nucleotide base sequence and cellular lipid composition. Strain F199 contained 2-OH14:0 and 18:1 omega 7c as the predominant cellular fatty acids and sphingolipids that are characteristic of the genus Sphingomonas. Phylogenetic analysis of its 16S rRNA sequence indicated that F199 was most closely related to Sphingomonas capsulata among the bacteria currently in the Ribosomal Database. Five additional isolates from deep Southeast Coastal Plain sediments were determined by 16S rRNA sequence analysis to be closely related to F199. These strains also contained characteristic sphingolipids. Four of these five strains could also grow on a broad range of aromatic compounds and could mineralize [14C]toluene and [14C]naphthalene. S. capsulata (ATCC 14666), Sphingomonas paucimobilis (ATCC 29837), and one of the subsurface isolates were unable to grow on any of the aromatic compounds or mineralize toluene or naphthalene. These results indicate that bacteria within the genus Sphingomonas are present in Southeast Coastal Plain subsurface sediments and that the capacity for degrading a broad range of substituted aromatic compounds appears to be common among Sphingomonas species from this environment.

Bacteria, Aerobic↗
PubMed1995

Single-molecule mechanics of heavy meromyosin and S1 interacting with rabbit or Drosophila actins using optical tweezers.

Single-molecule mechanical interactions between rabbit heavy meromyosin (HMM) or subfragment 1 (S1) and rabbit actin were measured with an optical tweezers piconewton, nanometer transducer. Similar intermittent interactions were observed with HMM and S1. The mean magnitude of the single interaction isotonic displacements was 20 nm for HMM and 15 nm with S1. The mean value of the force of single-molecule interactions was 1.8 pN for HMM and 1.7 pN with S1. The stiffness of myosin S1 was determined by applying a sinusoidal length change to the thin filament and measuring the corresponding force; the mean stiffness was 0.13 pN nm-1. By moving an actin filament over a long distance past an isolated S1 head, we found that cross-bridge attachment occurred preferentially at a periodicity of about 40 nm, similar to that of the actin helical repeat. Rate constants for the probability of detachment of HMM from actin were determined from histograms of the lifetime of the attached state. This gave a value of 8 s-1 or 0.8 x 10(6) M-1 s-1 for binding of ATP to the rigor complex. We conclude (1) that our HMM-actin interactions involve just one head, (2) that compliance of the cross-bridge is not in myosin subfragment 2, although we cannot say to what extent contributions arise from myosin S1 or actin, and (3) that the elemental movement can be caused by a change of shape of the S1 head, but that this would have to be much greater than the movements suggested from structural studies of S1 (Rayment et al., 1993).

Actins↗
PubMed1994

Comparison of phylogenetic relationships based on phospholipid fatty acid profiles and ribosomal RNA sequence similarities among dissimilatory sulfate-reducing bacteria.

Twenty-five isolates of dissimilatory sulfate-reducing bacteria were clustered based on similarity analysis of their phospholipid ester-linked fatty acids (PLFA). Of these, 22 showed that phylogenetic relationships based on the sequence similarity of their 16S rRNA directly paralleled the PLFA relationships. Desulfobacter latus and Desulfobacter curvatus grouped with the other Desulfobacter spp. by 16S rRNA comparison but not with the PLFA analysis as they contained significantly more monoenoic PLFA than the others. Similarly, Desulfovibrio africanus clustered with the Desulfovibrio spp. by 16S rRNA but not with them when analyzed by PLFA patterns because of higher monoenoic PLFA content. Otherwise, clustering obtained with either analysis was essentially congruent. The relationships defined by PLFA patterns appeared robust to shifts in nutrients and terminal electron acceptors. Additional analyses utilizing the lipopolysaccharide-lipid A hydroxy fatty acid patterns appeared not to shift the relationships based on PLFA significantly except when completely absent, as in Gram-positive bacteria. Phylogenetic relationships between isolates defined by 16S rRNA sequence divergence represent a selection clearly different from the multi-enzyme activities responsible for the PLFA patterns. Determination of bacterial relationships based on different selective pressures for various cellular components provides more clues to evolutionary history leading to a more rational nomenclature.

Desulfovibrio↗
PubMed1994

Optical biosensor for environmental on-line monitoring of naphthalene and salicylate bioavailability with an immobilized bioluminescent catabolic reporter bacterium.

An optical whole-cell biosensor based on a genetically engineered bioluminescent catabolic reporter bacterium was developed for continuous on-line monitoring of naphthalene and salicylate bioavailability and microbial catabolic activity potential in waste streams. The bioluminescent reporter bacterium, Pseudomonas fluorescens HK44, carries a transcriptional nahG-luxCDABE fusion for naphthalene and salicylate catabolism. Exposure to either compound resulted in inducible bioluminescence. The reporter culture was immobilized onto the surface of an optical light guide by using strontium alginate. This biosensor probe was then inserted into a measurement cell which simultaneously received the waste stream solution and a maintenance medium. Exposure under defined conditions to both naphthalene and salicylate resulted in a rapid increase in bioluminescence. The magnitude of the response and the response time were concentration dependent. Good reproducibility of the response was observed during repetitive perturbations with either naphthalene or salicylate. Exposure to other compounds, such as glucose and complex nutrient medium or toluene, resulted in either minor bioluminescence increases after significantly longer response times compared with naphthalene or no response, respectively. The environmental utility of the biosensor was tested by using real pollutant mixtures. A specific bioluminescence response was obtained after exposure to either an aqueous solution saturated with JP-4 jet fuel or an aqueous leachate from a manufactured-gas plant soil, since naphthalene was present in both pollutant mixtures.

Base Sequence↗
PubMed1994

Pseudomonas aeruginosa AlgG is a polymer level alginate C5-mannuronan epimerase.

Alginate is a viscous extracellular polymer produced by mucoid strains of Pseudomonas aeruginosa that cause chronic pulmonary infections in patients with cystic fibrosis. Alginate is polymerized from GDP-mannuronate to a linear polymer of beta-1-4-linked residues of D-mannuronate and its C5-epimer, L-guluronate. We previously identified a gene called algG in the alginate biosynthetic operon that is required for incorporation of L-guluronate residues into alginate. In this study, we tested the hypothesis that the product of algG is a C5-epimerase that directly converts D-mannuronate to L-guluronate. The DNA sequence of algG was determined, and an open reading frame encoding a protein (AlgG) of approximately 60 kDa was identified. The inferred amino terminus of AlgG protein contained a putative signal sequence of 35 amino acids. Expression of algG in Escherichia coli demonstrated both 60-kDa pre-AlgG and 55-kDa mature AlgG proteins, the latter of which was localized to the periplasm. An N-terminal analysis of AlgG showed that the signal sequence was removed in the mature form. Pulse-chase experiments in both E. coli and P. aeruginosa provided evidence for conversion of the 60- to the 55-kDa size in vivo. Expression of algG from a plasmid inan algG (i.e., polymannuronate-producing) mutant of P. aeruginosa restored production of an alginate containing L-guluronate residues. The observation that AlgG is apparently processed and exported from the cytoplasm suggested that it may act as a polymer-level mannuronan C5-epimerase. An in vitro assay for mannuronan C5 epimerization was developed wherein extracts of E. coli expressing high levels of AlgG were incubated with polymannuronate. Epimerization of D-mannuronate to L-guluronate residues in the polymer was detected enzymatically, using a L-guluronate-specific alginate lyase of Klebsiella aerogenes. Epimerization was also detected in the in vitro reaction between recombinant AlgG and poly-D-mannuronate, using high-performance anion-exchange chromatography. The epimerization reaction was detected only when acetyl groups were removed from the poly-D-mannuronate substrate, suggesting that AlgG epimerization activity in vivo may be sensitive to acetylation of the D-mannuronan residues. These results demonstrate that AlgG has polymer-level mannuronan C5-epimerase activity.

Alginates↗
PubMed1993

Detection of Legionella pneumophila in biofilms containing a complex microbial consortium by gas chromatography-mass spectrometry analysis of genus-specific hydroxy fatty acids.

A gas chromatographic-mass spectrometric method was used to detect Legionella pneumophila in biofilms in potable water containing a complex microbial consortium. The unique 3-hydroxy and 2,3-dihydroxy fatty acids of the L. pneumophila lipopolysaccharides (LPS) were detected in both the planktonic phase of the continuous culture model and in the biofilms forming on both copper and polyethylene substrata. The technique confirmed that lower numbers of Legionella colonised and grew on copper in comparison to polyethylene and offers promise for routine detection of Legionella in biofilms in the environment.

Fatty Acids↗
PubMed1993

Stimulation of secretory antibodies against Bordetella pertussis antigens in the lungs of mice after oral or intranasal administration of liposome-incorporated cell-surface antigens.

Lipopolysaccharide (LPS) and outer membrane protein (OMP) preparations of Bordetella pertussis were incorporated into multilamellar liposomes composed of soya bean-derived phospholipids which were then used for oral and intranasal immunization of mice. Specific antibody responses of animals immunized by either route were measured in lung washes. A specific IgA response to LPS was detected after immunization with the OMP-containing liposomes but not with the LPS-containing liposomes, indicating adjuvant activity of the proteins. The OMP-containing liposomes were significantly more effective in inducing immune responses than the OMP preparation alone. Responses were highest when mice were given a booster 30 days after primary immunization. Maximum responses occurred 20 days after the booster but specific antibody was still detected 75 days after the secondary immunization. These results suggest that this liposome antigen delivery system has potential in stimulating secretory antibody responses which may be helpful in protecting against infection from B. pertussis.

Administration, Intranasal↗
PubMed1993

Geobacter metallireducens gen. nov. sp. nov., a microorganism capable of coupling the complete oxidation of organic compounds to the reduction of iron and other metals.

The gram-negative metal-reducing microorganism, previously known as strain GS-15, was further characterized. This strict anaerobe oxidizes several short-chain fatty acids, alcohols, and monoaromatic compounds with Fe(III) as the sole electron acceptor. Furthermore, acetate is also oxidized with the reduction of Mn(IV), U(VI), and nitrate. In whole cell suspensions, the c-type cytochrome(s) of this organism was oxidized by physiological electron acceptors and also by gold, silver, mercury, and chromate. Menaquinone was recovered in concentrations comparable to those previously found in gram-negative sulfate reducers. Profiles of the phospholipid ester-linked fatty acids indicated that both the anaerobic desaturase and the branched pathways for fatty acid biosynthesis were operative. The organism contained three lipopolysaccharide hydroxy fatty acids which have not been previously reported in microorganisms, but have been observed in anaerobic freshwater sediments. The 16S rRNA sequence indicated that this organism belongs in the delta proteobacteria. Its closest known relative is Desulfuromonas acetoxidans. The name Geobacter metallireducens is proposed.

Cytochrome c Group↗
PubMed1993

Biodegradation of chlorinated aliphatics and aromatic compounds in total-recycle expanded-bed biofilm reactors.

Ground-water contamination by chlorinated aliphatic compounds is a major cause for concern because of their toxicity. This study examined the biodegradation of trichloroethylene and aromatic compounds by microbial consortia enriched from contaminated subsurface sediments. The consortia were capable of utilizing methane and propane as sources of carbon and energy. Two continuously recycled expanded-bed bioreactors were inoculated with (1) the subsurface consortium, and (2) P. fluorescence, P. putida (strains pRB1401 and pWWO), and M. trichosporium OB3b. An uninoculated reactor containing 0.2% sodium azide and 0.5% formalin served as the control. Methane (5% v/v) and propane (3% v/v) were maintained by batch feeding through the course of the experiment. Greater than 97% degradation of trichloroethylene was observed over a period of 12 d. More than 99% of benzene, toluene, and xylene were degraded within the first 7 d. Dissolved oxygen levels were measured and found to be in the range 4.9-6.5 mg/L throughout the experiments.

Benzene↗
PubMed1993

Biodegradation of chlorinated aliphatic hydrocarbon mixtures in a single-pass packed-bed reactor.

Aliphatic chlorinated compounds, such as trichloroethylene (TCE) and tetrachloroethylene (PCE), are major contaminants of ground water. A single-pass packed-bed bioreactor was utilized to study the biodegradation of organic waste mixtures consisting of PCE, TCE, and other short-chain chlorinated organics. The bioreactor consisted of two 1960-mL glass columns joined in a series. One column was packed with sand containing a microbial consortia enriched from a contaminated site. The other column provided a reservoir for oxygen and a carbon source of methane/propane that was recirculated through the reactor. Sampling was accomplished by both direct headspace and liquid effluent concentration analyses. The reactor was operated in a single-pass mode. Greater than 99% degradation of trichloroethylene, approaching drinking water standards, was observed when the bioreactor residence time ranged from 1.9 to 3.2 d. Typically, when the reactor was pulse-fed with methane, propane, and air, 1 mol of TCE was degraded/110 mol of substrate utilized. Perturbation studies were performed to characterize reactor behavior. The system's degradation behavior was affected by providing different carbon sources, a pulse feeding regime, supplementing microbial biomass, and by altering flow rates.

Bacteria↗
PubMed1993

Components of the inspiratory-arterial isoflurane partial pressure difference.

We have measured the partial pressure of isoflurane simultaneously in inspired gas (PIiso), end-expired gas (PE'iso), mixed-expired gas (PEiso), arterial (Paiso) and mixed venous blood (Pviso) in six patients (aged 57-79 yr) anaesthetized with nitrous oxide, oxygen and isoflurane before surgery and after PE'iso had been stable for at least 15 min. We related these changes to the various indices of pulmonary maldistribution to determine if they were sufficient to explain reported differences between PE'iso and Paiso. Alveolar deadspace dilution of end-expired gas was calculated for carbon dioxide and this dilution factor used to calculate the "ideal" alveolar Piso (PAiso) from the observed inspired and end-expired concentrations. Shunt fraction was measured for oxygen and then used to calculate the partial pressure of isoflurane in the pulmonary end-capillary blood (Pc'iso) from the partial pressure in arterial and mixed venous blood. Mean (SE) values were: PIiso 0.69 (0.05) kPa; PE'iso 0.52 (0.04) kPa; PAiso 0.50 (0.04) kPa; Pc'iso 0.38 (0.04) kPa; Paiso 0.35 (0.03) kPa and Pviso 0.22 (0.02) kPa; Paiso: PE'iso 0.66 (0.02) kPa. The mean "ideal" alveolar to pulmonary end-capillary Piso difference was 0.12 (0.01) kPa and highly significant (P < 0.001). Paiso was substantially less than PE'iso but, for isoflurane, the difference was reasonably constant (range 0.14-0.22 kPa). The difference was attributable in part to the effects of shunt and deadspace, but also a failure of equilibration of isoflurane between the alveolar gas and pulmonary end-capillary blood. It is likely to be different for other anaesthetics. We conclude that, while PE'iso may adequately reflect Paiso for isoflurane, it cannot be assumed that the relation between end-expiratory gas and arterial partial pressures is the same for all anaesthetics.

Aged↗
PubMed1993

Isolation and characterization of an N-methylcarbamate insecticide-degrading methylotrophic bacterium.

A gram-negative bacterium which hydrolyzed aryl N-methylcarbamate insecticides was isolated from an agricultural soil which quickly degraded these pesticides. This organism, designated strain ER2, grew on carbofuran as a sole source of carbon and nitrogen with a doubling time of 3 h in a mineral salts medium. The aromatic nucleus of the molecule was not metabolized, and carbofuran 7-phenol accumulated as the end product of metabolism. The insecticides carbaryl, bendiocarb, and propoxur were similarly hydrolyzed, with each yielding the corresponding phenol. Strain ER2 contained two plasmids (120 and 130 kb). A probe cloned from the pDL11 plasmid of Achromobacter sp. strain WM111, which encodes the carbofuran hydrolase (mcd) gene (P. H. Tomasek and J. S. Karns, J. Bacteriol. 171:4038-4044, 1989), hybridized to the 120-kb plasmid. Restriction fragment profiles of pDL11 and strain ER2 plasmid DNAs suggested that the 120-kb plasmid of strain ER2 is very similar to pDL11. On the basis of the results of biochemical tests, 16S rRNA sequence analysis, and membrane lipid analyses, strain ER2 was found to be a phylogenetically unique type II methylotroph. The constitutive carbofuran hydrolase activity in glucose-grown cells increased sevenfold when strain ER2 was grown in the presence of 100 mg of carbofuran per liter as the sole source of carbon and nitrogen or as the sole nitrogen source in the presence of glucose. Growth on carbofuran resulted in the induction of enzymes required for methylamine-dependent respiration and the serine pathway of formaldehyde assimilation. These results indicate that the carbofuran hydrolase mcd gene is conserved on a plasmid found in organisms from different geographic areas and that the specific activity of carbofuran degradation may increase in response to carbofuran treatment.

Alcaligenes↗
PubMed1993

Survival of subsurface microorganisms exposed to UV radiation and hydrogen peroxide.

Aerobic and microaerophilic subsurface bacteria were screened for resistance to UV light. Contrary to the hypothesis that subsurface bacteria should be sensitive to UV light, the organisms studied exhibited resistance levels as efficient as those of surface bacteria. A total of 31% of the aerobic subsurface isolates were UV resistant, compared with 26% of the surface soil bacteria that were tested. Several aerobic, gram-positive, pigmented, subsurface isolates exhibited greater resistance to UV light than all of the reference bacterial strains tested except Deinococcus radiodurans. None of the microaerophilic, gram-negative, nonpigmented, subsurface isolates were UV resistant; however, these isolates exhibited levels of sensitivity similar to those of the gram-negative reference bacteria Escherichia coli B and Pseudomonas fluorescens. Photoreactivation activity was detected in three subsurface isolates, and strain UV3 exhibited a more efficient mechanism than E. coli B. The peroxide resistance of four subsurface isolates was also examined. The aerobic subsurface bacteria resistant to UV light tolerated higher levels of H2O2 than the microaerophilic organisms. The conservation of DNA repair pathways in subsurface microorganisms may be important in maintaining DNA integrity and in protecting the organisms against chemical insults, such as oxygen radicals, during periods of slow growth.

Bacteria, Aerobic↗
PubMed1993

Phylogenetic analysis of a bacterial aerobic degrader of azo dyes.

Eubacterial consensus oligonucleotide primers were used to amplify by polymerase chain reaction the nearly full-length 16S rRNA gene of isolate C7, a gram-negative rod capable of aerobic degradation of azo dyes. The DNA product was cloned and sequenced. Phylogenetic analysis based upon this DNA sequence places C7 within the alpha subdivision of proteobacteria, most closely related to Caulobacter subvibrioides. The phospholipid fatty acid pattern resembles that of caulobacters, with monounsaturated 16- and 18-carbon fatty acids predominating. C7 is unusual in having a monounsaturated branched fatty acid in the phospholipids and exclusively 2-hydroxy fatty acids in the lipid-extracted residue. This organism is of potential use in bioreactors operated for azo dye degradation.

Aerobiosis↗
PubMed1991

Kinetics of ATP release and Pi binding during the ATPase cycle of lethocerus flight muscle fibres, using phosphate-water oxygen exchange.

Rate constants have been obtained using oxygen isotope exchange techniques for steps controlling ATP release and Pi binding in the ATPase cycle of insect flight muscle fibres from the giant waterbug Lethecerus. The new exchange data for Pi binding and ATP release are compatible with a model developed previously in which only the rate constants controlling Pi and ATP release change during fibre activation. Phosphate-water oxygen exchange occurs into ATP remaining after partial hydrolysis by chemically skinned fibres in (18O) water. For fully activated fibres, the results are compatible with a single set of rate constants controlling this exchange and give a rate constant for ATP release of 1 s-1 (21 degrees C, pH 7.0 I = 120 mM). Oxygen exchange also occurs between (18O4)Pi in the medium and water during ATP hydrolysis. There is a strong correlation between the measured rate constant of exchange and the value of keat for the ATPase activity at different levels of activation. For fibres fully activated by oscillation or strain, the rate constant for Pi binding to an actomyosin. ADP state is greater than 960 M-1 s-1.

Adenosine Diphosphate↗