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Biomedical subjects

W R Mayberry

Publications and source records attributed to W R Mayberry.

At least 55 records · Page 3Linked to original sources

Distribution and composition of lipopolysaccharides from mycoplasmas.

Polymeric carbohydrates containing glycerol and fatty acids were isolated from whole cells and membranes of mycoplasmas by hot aqueous phenol extraction and gel filtration. Lipopolysaccharides were found to occur in four species of Acholeplasma, two of Anaeroplasma, and in Mycoplasma neurolyticum. None were detected in Spiroplasma citri or in five species of Mycoplasma. All lipopolysaccharides contained both neutral and N-acylated amino sugars in ratios varying from 1:1 to 3:1. The neutral sugars found in varying distribution were glucose, galactose, and mannose. The amino sugars included fucosamine, an unidentified deoxyhexosamine, galactosamine, and glucosamine. Fucosamine and glucose were the only sugars common to all lipopolysaccharides. The fatty acids were similar to those found in the lipids of each organism.

Acholeplasma↗

Plasmalogen composition of Anaeroplasma.

The polar lipids of Anaeroplasma contained 33.1 percent alk-1'-enyl glyceryl ether (plasmalogen) form. Phosphatidylglycerol was the major polar lipid (55.2 percent) and contained nearly all of the plasmalogen. The alk-1'-enyl glyceryl ether form accounted for 58.3 percent of the phosphatidylglycerol.

Aldehydes↗

Immunological analysis of glycolipids and lipopolysaccharides derived from various mycoplasmas.

The lipids and lipopolysaccharides of five mycoplasmas were examined for complement-fixing activity to antimembrane rabbit sera. Total glycolipid fractions and the aqueous phenol fractions (lipopolysaccharides) from the membranes of Acholeplasma laidlawii, A. modicum, A. axanthum, and Mycoplasma neurolyticum exhibited significant antigenic activity. The glycolipids and phosphoglycolipids from Thermoplasma acidophilum were either anticomplementary or did not react due to their extreme hydrophobic nature. High activity was found using the lipopolysaccharide of T. acidophilum. The pure glycolipids and phosphoglycolipids of the Acholeplasma and Mycoplasma species also exhibited significant complement-fixing activity. Acylation of the sugar residues of these lipids reduced or negated complement-fixing activity. Double cross-reactions between the glycolipids of A. laidlawii and A. modicum appeared to be due to mono- and diglucosyl diglycerides of identical structure. Specificity of glycolipid structure was noted by the absence of cross-reactions between A. laidlawii and M. neurolyticum, the glycolipids of which differ only in the nature of the glucose linkages. The existence of lipopolysaccharides in the membranes of mycoplasmas and their complement-fixing activity in the presence of antimembrane sera suggest their possible importance as specific antigenic determinants.

Animals↗

Heptose-containing pentaglycosyl diglyceride among the lipids of Acholeplasma modicum.

A pentaglycosyl diglyceride with the tentative structure of galactosyl-galactosyl-mannoheptosyl-glucosyl-glucosyl diglyceride was found to be the major glycolipid in Acholeplasma modicum. The heptose is d-glycero-d-mannoheputose. The diglyceride-terminating moiety possesses the structure O-alpha-d-glucopyranosyl-(1 --> 2)-O-alpha-d-glucopyranosyl-sn-1,2-diglyceride. Other glycolipids occurring in this organism are a diglucosyl diglyceride and a monoglucosyl diglyceride with structures identical to the terminal segments of the pentaglycosyl diglyceride. More fully acylated derivatives of these two glycolipids also occur. The phospholipids are all of the glycerophosphoryl type. The neutral lipids are composed of diglycerides and four polyterpenes. The polyterpenes consist of both colored and colorless carotenoids and become radiolabeled with both [(14)C]acetate and [(14)C]mevalonate.

Acetates↗

Long-chain glycerol diether and polyol dialkyl glycerol triether lipids of Sulfolobus acidocaldarius.

Cells of Sulfolobus acidocaldarius contain about 2.5% total lipid on a dry-weight basis. Total lipid was found to contain 10.5% neutral lipid, 67.6% glycolipid, and 21.7% polar lipid. The lipids contained C(40)H(80) isopranol glycerol diethers. Almost no fatty acids were present. The glycolipids were composed of about equal amounts of the glycerol diether analogue of glucosyl galactosyl diglyceride and a glucosyl polyol glycerol diether. The latter compound contained an unidentified polyol attached by an ether bond to the glycerol diether. The polar lipids contained a small amount of sulfolipid, which appeared to be the monosulfate derivative of glucosyl polyol glycerol diether. About 40% of the lipid phosphorus was found in the diether analogue of phosphatidyl inositol. The remaining lipid phosphorus was accounted for by approximately equal amounts of two inositol monophosphate-containing phosphoglycolipids, inositolphosphoryl glucosyl galactosyl glycerol diether and inositolphosphoryl glucosyl polyol glycerol diether.

Autoradiography↗

Characterization of the membranes of Thermoplasma acidophilum.

Thermoplasma acidophilum grows optimally under aeration at 59 C and pH 2. Both intact cells and membranes disaggregate below pH 1 and above pH 5, producing no sedimentable particles. Increase in ionic strength at pH 5 or below results in cellular lysis and membrane disaggregation. Membranous components produced by lysis at alkaline pH reaggregate upon reduction of both pH and ionic strength. Osmotic environment plays little role in cellular stability. Membranes prepared by sonic lysis at pH 5 exhibit vesicular structures and are composed of multiple proteins. Although the amino acid composition of the membrane proteins is similar to other mycoplasmal membranes, the number of free amino and carboxyl groups is less than half of those in Acholeplasma. Reduction of the number of free carboxyl groups results in membrane stabilization over a wide range of pH. Increase in the number of free amino groups reverses the stability of membranes relative to pH. Acidophily in Thermoplasma can be related to a significant reduction in repulsing negative charges on the membrane proteins.

Amino Acids↗

Identification of the amide-linked fatty acids of Acholeplasma axanthum S743 as D(-)3-hydroxyhexadecanoate and its homologues.

The amide-linked fatty acids of the sphingolipids of Acholeplasma axanthum S743 are predominantly hydroxy acids. These acids were shown by gas-liquid chromatography, mass spectrometry, and polarimetry to be the d(-)3-hydroxy fatty acids. The predominant component of the mixture was 3-hydroxyhexadecanoate (beta-hydroxypalmitate, hydroxy [h] 16:0) followed by h 20Delta (Delta = unsaturated), h14:0, h12:0, and h18Delta in decreasing order of concentration. The fatty acid profile indicates that these beta-hydroxy acids possibly arise from elongation of the fatty acids supplied in the growth medium.

Bacteria↗

Lipids of a T strain of Mycoplasma.

Cholesterol, free fatty acids, and phosphatidic acid are the predominant lipids of a T strain of Mycoplasma. The remaining neutral lipids are composed of cholesteryl esters, triglycerides, and diglycerides. Three glucose-containing glycolipids are present in trace amounts. In addition to phosphatidic acid, the phospholipids are comprised of phosphatidyl glycerol, diphosphatidyl glycerol, and phosphatidyl ethanolamine. Another polar lipid was found to be ninhydrin-positive and phosphate-free. It appears to be a diamino hydroxy compound containing adjacent fatty acid ester and N-acyl groups.

Autoradiography↗

Lipids of Thermoplasma acidophilum.

Cells of Thermoplasma acidophilum contain about 3% total lipid on a dry weight basis. Total lipid was found to contain 17.5% neutral lipid, 25.1% glycolipid, and 56.6% phospholipid by chromatography on silicic acid. The lipids contain almost no fatty acid ester groups but appear to have long-chain alkyl groups in ether linkages to glycerol. The phospholipid fraction includes a major component which represents about 80% of the lipid phosphorus and 46% of the total lipids. We believe this component to be a long-chain isopranol glycerol diether analogue of glycerolphosphoryl monoglycosyl diglyceride. The glycolipids appear to contain isopranol diether analogues. Several components of the complex, neutral lipid fraction have been identified as hydrocarbons, vitamin K(2)-7, and isopranol glycerol diether analogues. Sterols are present in the neutral lipids but do not appear to be synthesized by the organism.

Autoradiography↗

Calorific content of certain bacteria and fungi.

Calorific contents of dried cells of several representative species of bacteria (gram-negative rods and gram-positive rods and cocci), two species of yeasts, and a filamentous fungus were determined by bomb calorimetry. The grand mean was 5,383 cal per g of ash-free dry weight. This value was then used to determine quantity of energy assimilated (E(a)) during growth. Subsequently, E(a) was employed in the equation: Y(kcal) = Y/(E(a) + E(d)), where Y(kcal) is the yield of cells per kilocalorie of energy taken from a culture medium, Y is the yield per mole of substrate utilized, E(a) is Y times caloric content of the cells, and E(d) is the energy expended by oxidative dissimilation. An estimate of E(d) was obtained for a number of experiments by multiplying the moles of oxygen consumed per mole of substrate utilized during growth by the average quantity of energy utilized to reduce a mole of oxygen with electrons from organic compounds (106 kcal). From previous studies in our laboratories, a value for Y(kcal) of 0.118 g/kcal was predicted. The mean value for data from five studies of aerobic growth of prototrophic heterotrophs was found to be 0.111.

Air↗

Lipids of a sterol-nonrequiring Mycoplasma.

The lipids of the sterol nonrequiring Mycoplasma strain S743 were found to include both ester glycerophosphatides (phosphatidylglycerol, acylphosphatidylglycerol, and diphosphatidylglycerol) and ceramide glycerophosphate compounds containing N-hydroxyacyl groups. The major phosphosphingolipid was tentatively identified as a hydroxyceramidephosphorylglycerol containing an O-acyl group. These compounds became labeled during growth in the presence of (32)P-orthophosphate, (14)C-glycerol, or (14)C-palmitate. The lipid fraction also contained free long-chain base. (14)C-palmitate was converted to labeled sphinganine. The long-chain base composition of the lipids was modified by growing the organisms in media containing different fatty acids, which were converted to bases containing two more C atoms per molecule. Ninety per cent of the long-chain base from cells grown in medium supplemented with elaidate consisted of monounsaturated C(20) base.

Autoradiography↗

Observations on membranes of Mycoplasma laidlawii strain B.

The cytoplasmic membrane of Mycoplasma laidlawii strain B is solubilized by anionic and nonionic detergents, succinylation, phospholipase A, alkaline phosphatase, trypsin, and chymotrypsin. Cationic detergents are without effect, as are chelating agents, even in the presence of high concentrations of monovalent cation. The detergent-solubilized membrane exhibits one peak in the analytical ultracentrifuge, but the sedimentation coefficient is dependent upon concentration of detergent. Simple dialysis does not remove all of the sodium dodecylsulfate except from lipid-depleted membrane particles. Membranes bind sodium dodecylsulfate but acetone powders of membranes do not. Sulfated alcohols with chain lengths of C(14) and C(16) are more tightly bound than dodecylsulfate. A constant amount of di- and trivalent cation is bound by the membrane upon aggregation. Only a portion of this cation is removable with chelating agents. No chelating agent is bound by these aggregates. A portion of the lipid-depleted membrane particles is solubilized by negatively charged lipids and detergents, giving rise to aggregates in the presence of divalent cation. Fractionations of detergent-solubilized membranes by preparative gel electrophoresis and ammonium sulfate were inconclusive. Density gradient centrifugation of succinylated membranes yielded at least five fractions which exhibited homogeneity by ultracentrifugation. Analytical gel electrophoresis of these fractions demonstrated heterogeneity. The composition of these five fractions suggested separation of protein from lipid.

Alkaline Phosphatase↗

Growth yields of bacteria on selected organic compounds.

Cell yields were determined for two bacterial soil isolants grown aerobically in minimal media on a variety of synthetic organic compounds. 1-Dodecanol, benzoic acid, phenylacetic acid, phenylglyoxylic acid, and diethylene, triethylene, and tetraethylene glycols were tested. Two "biochemicals," succinate and acetate, were also tested for comparison. Yields were calculated on the basis of grams of cells obtained per mole of substrate utilized, gram atom of carbon utilized, mole of oxygen consumed, and equivalent of "available electrons" in the substrates. This latter value appears to be nearly constant at 3 g of cells per equivalent of "available electrons." Yields predicted on this basis for other bacteria and for yeasts on other substrates are in fair agreement with reported values.

Journal Article↗

Metabolism of linear alcohols with various chain lengths by a Pseudomonas species.

Pseudomonas C12B grew on and oxidized linear primary alcohols with even- and odd-numbered carbon chains ranging from C(2) to C(11). Cell-free extracts of the bacteria contained a nicotinamide adenine dinucleotide-linked dehydrogenase(s) active with these alcohols and with branched primary and linear secondary alcohols as well. Analysis by gas-liquid chromatography of hexane extracts of filtrates of cultures containing mixtures of even-carbon numbered alcohols from C(10) to C(18) revealed that decanol was rapidly utilized, whereas the remainder were slowly dissimilated up to 19 hr and then were rapidly degraded in the next few hours of culture. The validity for these studies of (i) steam distillation as a method for collecting the alcohols from cultures, and (ii) quantitative estimation by gas-liquid chromatographic comparison with an added internal marker, was established. Steam distillation and gas-liquid chromatography were then used to show that failure to demonstrate stoichiometry of sulfate and dodecanol in the alkyl sulfatase reaction in a previous study resulted from contamination of the commercial "Dodecyl Sodium Sulfate, 95%" used with decyl, undecyl, and tetradecyl sulfates.

Alcohols↗

Primary alcohol sulfatase in a Pseudomonas species.

An ammonium sulfate-precipitated fraction from cell-free extracts of Pseudomonas C12B grown on a medium containing sodium dodecyl sulfate (SDS) contained alkyl sulfatase increased fourfold in specific activity over the crude. Optimal pH (7.5) and temperature (70 C) for sulfate release were determined with SDS labeled with radioactive sulfur (SDS(35)) as test substrate. Phosphate, arsenate, and certain heavy metal ions inhibited desulfation, whereas Mg(++) and Mn(++) stimulated activity of preparations which had been dialyzed against ethylenediaminetetraacetic acid. Dodecanol was recovered in semiquantitative yield from reaction mixtures containing enzyme and SDS(35). Aryl sulfates, secondary alcohol sulfates, and a phenoxyethyl sulfate failed to serve as substrate for this enzyme.

Alcohols↗