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Biomedical subjects

D C White

Publications and source records attributed to D C White.

At least 217 records · Page 12Linked to original sources
PubMed1971

Effect of growth temperature on the lipid composition of Thermus aquaticus.

The complex lipids of Thermus aquaticus include phospholipids, glucolipids, carotenoids, and vitamin K(2) isoprenologues. The phospholipids account for 30% of the total lipids and have been identified as phosphatidylethanolamine (4%), phosphatidylglycerol (3%), phosphatidylinositol (10%), cardiolipin (3%), and phosphatidic acid (1%). The major phospholipid contained three fatty acids, a long-chain unsaturated amine, and one glycerol per phosphate and accounted for 80% of the lipid phosphate. The carotenoids accounted for 60% of the membrane lipid. The majority of the carotenoids were very polar. Mono- and diglucosyldiglyceride and the 35-, 40-, and 45-carbon vitamin K(2) isoprenologues were also identified. All these lipids were localized in the membrane of T. aquaticus. When the growth temperature was increased from 50 to 75 C and measured at 5 C intervals, there was a progressive increase in the total lipid content. The phospholipids increased 2-fold, the carotenoids increased 1.8-fold, and the glucolipids increased 4-fold between cells grown at 50 C and 75 C. The vitamin K(2) level did not change. The proportions of the individual lipids within each lipid class remained constant as the temperature of growth was raised. Metabolic studies indicated turnover of the diacyl phospholipids during pulse-chase experiments at rates comparable with mesophilic bacteria. The major phospholipid and the carotenoids did not turn over.

Acetone↗
PubMed1971

Detection of a rapidly metabolizing portion of the membrane cardiolipin in Haemophilus parainfluenzae.

Heterogeneity in the metabolism of cardiolipin (CL) has been detected in Haemophilus parainfluenzae. Pulse-chase experiments showed that a portion of the total CL incorporated and then lost (32)P much more rapidly than the rest of the CL in the cells. The metabolism of each phosphate of the CL differed. The phosphate of the phosphatidyl glycerol (PG) portion of the CL had a more active metabolism than the phosphate of the phosphatidic acid portion of the molecule. Only a portion of the PG pool contributed to the formation of CL. Ethylenediaminetetraacetic acid inhibited the CL-specific phospholipase D in vitro and, when added to growing cells, resulted in more rapid PG metabolism, suggesting that CL hydrolysis contributed to the PG pool.

Autoradiography↗
PubMed1971

Consequences of the inhibition of cardiolipin metabolism in Haemophilus parainfluenzae.

Examination of phospholipid metabolism in Haemophilus parainfluenzae with inhibitors of various cellular functions indicated that macromolecular synthesis and lipid metabolism can be dissociated at least for a short time. Two classes of inhibitors have relatively specific effects on cardiolipin (CL) metabolism. Pentachlorophenol and p-hydroxymercuribenzoate blocked CL synthesis but allowed CL hydrolysis to phosphatidic acid and phosphatidyl glycerol (PG); 3,3',4,5'-tetrachlorosalicylanilide (TCS) and carbonyl cyanide m-chlorophenylhydrazone (m-CCCP) blocked CL hydrolysis with the stoichiometric accumulation of CL. It appeared as if TCS and m-CCCP inhibited a vital activity coupled with the hydrolysis of CL by the highly active, CL-specific phospholipase D found in this organism. Because TCS and m-CCCP are thought to act by destroying the proton gradient thereby interrupting energy-dependent transport, it is possible that a highly active portion of the cellular CL could be coupled to some phase of this process.

Anti-Bacterial Agents↗
PubMed1970

Rigor contraction and the effect of various phosphate compounds on glycerinated insect flight and vertebrate muscle.

1. Glycerol-extracted flight muscle from the giant water-bug, Lethocerus cordofanus, undergoes contraction when deprived of ATP. This rigor contraction occurs in the presence of the calcium chelating agent EGTA, under conditions in which the predicted free calcium ion concentration is less than 5 x 10(-9)M. A mechanically similar contraction has been observed by depriving the muscle of Mg(2+), in the presence of 5 mM-ATP.2. In the rigor contraction tensions of up to 120 mN/mm(2) under isometric conditions, and shortenings of between 2(1/2) and 6% under isotonic conditions, have been observed at 20 degrees C.3. Muscles in rigor can be relaxed by the addition of ATP. The minimum concentration of ATP required to give full relaxation depends upon other ionic constituents of the solutions, and upon temperature, but is between 0.3 and 1 mM at 20 degrees C. Lower concentrations result in partial reduction of tension and stiffness.4. Pyrophosphate (PP) causes a reduction in the tension of muscle which has developed rigor under isometric conditions, but the stiffness, when measured at frequencies between 1 and 100 Hz, remains indistinguishable from that of rigor muscle. The stiffness when measured by applying slow length changes is comparable to that of ATP-relaxed muscle. It is suggested that in PP-relaxed muscle the cross-bridges remain in close proximity to, but are not rigidly attached to, the I filaments, resulting in a high viscous interaction between the two sets of filaments.5. The addition of low concentrations of ADP (less than about 0.7 mM) to rigor muscle in the absence of ATP causes an increase in tension. The effect is sigmoid, and is probably due to the formation of low concentrations of ATP throughout the body of the fibre by myokinase activity. Larger concentrations of ADP (about 5 mM) added to rigor muscle cause relaxation, probably due to the formation of higher ATP concentrations.6. AMP and inorganic orthophosphate have little effect upon the mechanical properties of rigor muscle.7. There is a delay of about 1 min before the onset of rigor contraction when fibres are transferred from an ATP-solution to one containing no ATP, due to the transfer of ATP in the body of the fibres. Both ATP hydrolysis by the fibres and diffusion of ATP into the bathing solution contribute significantly to the rate of depletion of ATP.8. Rabbit psoas muscle shows a similar rigor contraction in the presence of EGTA, and has mechanical properties similar to those described for Lethocerus flight muscle in the presence of pyrophosphate.

Adenine Nucleotides↗
PubMed1970

Lipids of Bacteroides melaninogenicus.

The lipids of Bacteroides melaninogenicus were readily extractable with chloroform-methanol. Three per cent of the fatty acids were not extractable. The neutral lipids contained 4% of the extractable fatty acids, the stench characteristic of these organisms, and 0.5 mumole of vitamin K(2) isoprenologues K(2)-35, K(2)-40, and K(2)-45 per g (dry weight). This is one-fifth to one-tenth of the vitamin K(2) level found in other bacteria. Ninety-six per cent of the extractable fatty acids were associated with the phospholipids (60 mumoles of lipid phosphate/g, dry weight), which consisted of the diacyl lipids phosphatidic acid, phosphatidyl serine, and phosphatidyl ethanolamine (with phosphatidyl glycerol and cardiolipin in one strain). The unusual phosphosphingolipids ceramide phosphorylethanolamine, ceramide phosphorylglycerol, and ceramide phosphorylglycerol phosphate accounted for 50 to 70% of the lipid phosphate. In protoheme-requiring strains, the protoheme concentration in the growth medium regulated the growth rate and the amount of enzymatically reducible cytochrome c. There were no gross changes in the lipid composition in cells containing different levels of enzymatically reducible cytochrome c.

Bacteroides↗
PubMed1970

Effect of nitrate, fumarate, and oxygen on the formation of the membrane-bound electron transport system of Haemophilus parainfluenzae.

The composition of the membrane-bound electron transport system of Haemophilus parainfluenzae underwent modification in response to the terminal electron acceptor in the growth medium. H. parainfluenzae was able to grow with O(2), nitrate, fumarate, pyruvate, and substrate amounts of nicotinamide adenine dinucleotide (NAD) as electron acceptors. When O(2) served as the electron acceptor and its concentration was lowered below 20 mum, the bacteria formed more cytochromes b, c, a(1), a(2), and o than were present in the cells grown at 150 to 200 mum O(2). Nitrate and nitrite reductase activities also appeared during growth at the low O(2) concentrations in the absence of added nitrate. Cytochrome levels in cells grown anaerobically with fumarate, pyruvate, or NAD as terminal acceptors were similar to those formed in cells grown at low O(2) concentrations. Cells grown with nitrate had higher levels of cytochromes c, b, and o, and of nitrate and nitrite reductases, than did cells grown with the other acceptors. The formation of cytochrome oxidase a(2) was repressed by the presence of nitrate in the growth medium. The critical O(2) concentration (the O(2) concentration at which the rate of O(2) uptake becomes demonstrably dependent on the O(2) concentration) was about 100 mum in cells grown with nitrate and about 15 mum in cells grown with the other acceptors. A mutant of H. parainfluenzae was found to make about 10% as much cytochrome c as the wild type, and its formation of cytochrome a(2) was not repressed by nitrate. The critical O(2) concentration of the mutant was high when it was grown with nitrate, suggesting that the high levels of cytochrome c and the absence of cytochrome a(2) from the wild type are not responsible for the high critical O(2) concentration. The modifications of the respiratory system induced by changing the terminal electron acceptor were inhibited by the presence of chloramphenicol, which suggests that protein synthesis is involved.

Cell Membrane↗
PubMed1970

Release of membrane components from viable Haemophilus parainfluenzae by ethylenediaminetetraacetic acid-tris(hydroxymethyl)-aminomethane.

Logarithmically growing Haemophilus parainfluenzae lost 15 to 20% of the phospholipids, demethyl vitamin K(2), cytochrome b, and cytochrome c, and 50% of the lipopolysaccharide when incubated in ethylenediaminetetraacetic acid (EDTA)-tris-(hydroxymethyl)aminomethane (Tris) for 10 min. This loss of membrane components occurred without loss in viability, and the lost components were recovered as membrane fragments in the surrounding buffer. The phospholipids recovered in the membrane fragments had a slightly lower specific activity than the phospholipids in the residue. Lysis of a portion of the cells could not account for the release of membrane components, as the cells lost neither glucose-6-phosphate dehydrogenase activity not deoxyribonucleic acid. The treated cells were osmotically stable and contained the same proportions of the individual phospholipids as pretreatment cells. Prolongation of the EDTA-Tris treatment did not induce further loss of phospholipid or demethyl vitamin K(2), but caused a decrease in viability. If the cells were returned to the growth medium after 10 min, the cells immediately resumed growth at the pretreatment rate. During growth in the recovery period, the phospholipids increased logarithmically in the pretreatment rate. During growth in the recovery period, the phospholipids increased logarithmically in the pretreatment proportions, although there was a marked decrease in the turnover and a shift from the use of extracellular lipid precursors to the use of intracellular pools of precursors.

Buffers↗
PubMed1970

Heterogeneity of phospholipid composition in the bacterial membrane.

Heterogeneity in the distribution or binding of the membrane phospholipids was demonstrated in the membrane fragments released from Haemophilus parainfluenzae by treatment with ethylenediaminetetraacetic acid (EDTA)-tris(hydroxymethyl)-aminomethane (Tris). The membrane fragments released early in the EDTA-Tris treatment contained two- to fivefold higher proportions of cardiolipin and phosphatidylglycerol and less phosphatidylethanolamine as well as phospholipids with threefold lower specific activity of the phospholipid phosphate after a short pulse of (32)P than were found in the residue. Heterogeneity was best demonstrated with shorter EDTA-Tris treatments and shorter periods of growth with (32)P. EDTA-Tris treatment appeared to progressively strip phospholipids from the cells that were synthesized at progressively later times.

Autoradiography↗
PubMed1970

Cardiolipin-specific phospholipase D activity in Haemophilus parainfluenzae.

A highly active phospholipase D that is specific for cardiolipin was detected in the gram-negative bacterium Haemophilus parainfluenzae. Previously reported phospholipase D preparations have come exclusively from higher plants. The bacterial enzyme hydrolyzed cardiolipin to phosphatidyl glycerol and phosphatidic acid. During the incubation, phosphatidic acid disappeared. Phosphatidyl ethanolamine, methylated phosphatidyl ethanolamines, phosphatidyl choline, and phosphatidyl glycerol were not hydrolyzed when cardiolipin was rapidly hydrolyzed.

Autoradiography↗
PubMed1970

Carotenoid formation by Staphylococcus aureus.

The carotenoid pigments of Staphylococcus aureus U-71 were identified as phytoene; zeta-carotene; delta-carotene; phytofluenol; a phytofluenol-like carotenoid, rubixanthin; and three rubixanthin-like carotenoids after extraction, saponification, chromatographic separation, and determination of their absorption spectra. There was no evidence of carotenoid esters or glycoside ethers in the extract before saponification. During the aerobic growth cycle the total carotenoids increased from 45 to 1,000 nmoles per g (dry weight), with the greatest increases in the polar, hydroxylated carotenoids. During the anaerobic growth cycle, the total carotenoids increased from 20 nmoles per g (dry weight) to 80 nmoles per g (dry weight), and only traces of the polar carotenoids were formed. Light had no effect on carotenoid synthesis. About 0.14% of the mevalonate-2-(14)C added to the culture was incorporated into the carotenoids during each bacterial doubling. The total carotenoids did not lose radioactivity when grown in the absence of (14)C for 2.5 bacterial doublings. The total carotenoids did not lose radioactivity when grown in the absence of (14)C for 2.5 bacterial doublings. The incorporation and turnover of (14)C indicated the carotenes were sequentially desaturated and hydroxylated to form the polar carotenoids.

Autoradiography↗
PubMed1970

Metabolism of phospholipid 2-linked fatty acids during the release of membrane fragments from Haemophilus parainfluenzae by ethylenediaminetetraacetic acid-tris(hydroxymethyl)-aminomethane.

Membrane fragments containing diacyl phospholipids were released from viable cells of Haemophilus parainfluenzae during incubation in ethylenediaminetetraacetic acid (EDTA)- tris(hydroxymethyl)aminomethane (Tris) buffer. The phospholipids located in the part of the membrane that was released during the EDTA-Tris treatment had markedly different proportions of fatty acids than the lipids remaining in the cell residue. Very little metabolism of the 1-linked fatty acid occurred. After a short pulse with (14)C, the specific activity of the 1-linked fatty acid was lower in the phospholipids released than in the phospholipids of the residue, indicating an earlier time of synthesis of those lipids released in the membrane fragments. During the EDTA-Tris treatment, the 2-linked fatty acid was metabolized. This metabolism may have involved phospholipase A(2) which stimulates the synthesis of fatty acids and the transfer of acyl groups to the lysophospholipid.

Acyltransferases↗
PubMed1970

Inhibition of carotenoid hydroxylation in Staphylococcus aureus by mixed-function oxidase inhibitors.

Compounds known to be inhibitors of mixed-function oxidase systems inhibited the aerobic synthesis of hydroxylated carotenoids in Staphylococcus aureus U-71. Growth of the cells in the presence of 2-diethylaminoethyl-2,2-diphenyl valerate, 2,4-dichloro-6-phenylphenoxyethylamine, 2,4-dichloro-6-phenylphenoxyethyldiethylamine, and piperonyl butoxide reduced the levels of the rubixanthins found in stationary-phase cells by 75 to 97%. In cells grown with mevalonate-2-(14)C, the turnover rate of phytoene was reduced and the turnover rate of phytofluenol was increased in the presence of these inhibitors. The zeta- and delta-carotenes, which turn over in the absence of the inhibitors, accumulated (14)C in the presence of the inhibitors. This suggested that a mixed-function oxidase was responsible for the aerobic hydroxylation of delta-carotene in S. aureus U-71.

Carbon Isotopes↗
PubMed1970

Inhibition of vitamin K2 and carotenoid synthesis in Staphylococcus aureus by diphenylamine.

Diphenylamine at concentrations which did not effect the growth rate inhibited the synthesis of vitamin K(2) in both anaerobic and aerobic cultures by about 50%. At this concentration, diphenylamine inhibited the synthesis of the cyclic carotenoids delta-carotene and the rubixanthins 25 to 35% anaerobically and 60 to 90% aerobically. The inhibition of synthesis of cyclic carotenoids and vitamin K(2) by diphenylamine had no detectable effect on the formation of the membrane-bound electron transport system.

Aniline Compounds↗
PubMed1970

Metabolism of the glycosyl diglycerides and phosphatidylglucose of Staphylococcus aureus.

A glucose containing lipid, phosphatidylglucose (probably 3-sn-phosphatidyl-1'-glucose) and a lipid tentatively identified as phosphatidylethanolamine have been characterized in the lipids of Staphylococcus aureus. These lipids together comprise less than 2% of the total phospholipids of exponentially growing S. aureus and accumulate to 14% of the total phospholipid in stationary-phase cells. These lipids lost no (32)P when cells grown with H(3) (32)PO(4) were transferred to nonradioactive medium during the exponential growth phase. This was in marked contrast to the other phospholipids which lost (32)P rapidly. The loss of (32)P from phosphatidic acid and cardiolipin in exponentially growing cells was biphasic, suggesting heterogeneity of phospholipid phosphate metabolism. The mono- and diglucosyl diglycerides showed a rapid loss of (14)C-glucose during growth in nonradioactive medium but no loss of (14)C from the fatty acids of these lipids. The (14)C in the glucose and fatty acids of the glucosyl diglycerides was derived from glucose.

Autoradiography↗