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Biomedical subjects

H U Koch

Publications and source records attributed to H U Koch.

At least 37 records · Page 2Linked to original sources

Maintenance of D-alanine ester substitution of lipoteichoic acid by reesterification in Staphylococcus aureus.

Toluene-treated Staphylococcus aureus cells did not synthesize teichoic acid and lipoteichoic acid under the conditions used. The organism displayed, however, a high capacity of incorporating D-[14C]alanine into previously formed polymers. The reaction was dependent on ATP and enhanced by magnesium ions. The incorporation rate into lipoteichoic acid correlated with the rate of loss of alanine ester which occurred through transfer to teichoic acid and base-catalyzed hydrolysis. At pH 6.5 the loss (20% within 4 h) was completely compensated for by reesterification. At pH 7.5 the loss was 60%, but by accelerated incorporation it was reduced to 10%. Incorporation was also enhanced when the original substitution of lipoteichoic acid was lowered by previous growth of S. aureus at high salt concentration. The newly added alanine was randomly distributed along the poly(glycerophosphate) chain. The decreased alanine substitution of lipoteichoic acid after growth at high salt concentration was shown to result from a direct inhibition of alanine incorporation.

Adenosine Triphosphate↗

The role of lipoteichoic acid biosynthesis in membrane lipid metabolism of growing Staphylococcus aureus.

Pulse-chase experiments with [2-3H]glycerol and [14C]acetate revealed that in Staphylococcus aureus lipoteichoic acid biosynthesis plays a dominant role in membrane lipid metabolism. In the chase, 90% of the glycerophosphate moiety of phosphatidylglycerol was incorporated into the polymer: 25 phosphatidylglycerol + diglucosyldiacylglycerol leads to (glycerophospho)25-diglucosyldiacylglycerol + 25 diacylglycerol. Glycerophosphodiglucosyldiacylglycerol was shown to be an intermediate, confirming that the hydrophilic chain is polymerized on the final lipid anchor. Total phosphatidylglycerol served as the precursor pool and was estimated to turn over more than twice for lipoteichoic acid synthesis in one bacterial doubling. Of the resulting diacylglycerol approximately 10% was used for the synthesis of glycolipids and the lipid anchor of lipoteichoic acid. The majority of diacylglycerol recycled via phosphatidic acid to phosphatidylglycerol. Synthesis of bisphosphatidylglycerol was negligible and only a minor fraction of phosphatidylglycerol passed through the metabolically labile lysyl derivative. In contrast to normal growth, energy deprivation caused an immediate switch-over from the synthesis of lipoteichoic acid to the synthesis of bisphosphatidylglycerol.

Diglycerides↗

Improved preparation of lipoteichoic acids.

A procedure is described for measuring the extraction of lipoteichoic acids from gram-positive bacteria in absolute terms. Virtually complete extraction was achieved from various bacteria by hot phenol/water if the cells were disrupted. Extraction of whole and delipidated cells and of the membrane fraction gave considerably lower yields. Most of the nucleic acids co-extracted from disrupted cells was removed by treatment with nucleases. Nuclease-resistant nucleic acid, protein, polysaccharide, and teichoic acid were separated from lipoteichoic acid by anionexchange chromatography on DEAE-Sephacel or hydrophobic interaction chromatography on octyl-Sepharose. Purified preparations were essentially free of polymeric contaminants, retained their alanine ester substitution, and were in the sodium salt form. Hydrophobic interaction chromatography also made it possible to recognize contamination of lipoteichoic acid with its deacylated and lyso-form, and to discriminate molecular species containing two and three, or two and four acyl groups.

Acetylation↗

Effect of alanine ester substitution and other structural features of lipoteichoic acids on their inhibitory activity against autolysins of Staphylococcus aureus.

Native substitution with the D-alanine ester of lipoteichoic acids (LTAs) affects their immunological properties, the capacity to bind divalent cations, and LTA carrier activity. In this study we tested the influence of the D-alanine ester on anti-autolytic activity, using extracellular autolysin from Staphylococcus aureus and nine LTAs with alanine/phosphorus molar ratios of between 0.23 and 0.71. The inhibitory activity, highest with alanine-free LTA, exponentially decreased with increasing alanine content, approaching zero at substitutions of greater than 0.6. Correspondingly, dipolar ionic phospholipids were not inhibitory, in contrast to negatively charged ones. Glycosylation of LTA up to an extent of 0.5 did not depress inhibitory activity, and even at a degree of 0.8 the effect was comparatively small. On comparison of LTAs from various sources, differences in lipid structures and chain lengths were without effect. The inhibitory activity drastically decreased when the glycolipid carried a single glycerophosphate residue or the hydrophilic chain had the unusual structure [6 leads to Gal(alpha 1--6)Gal(alpha 1--3)Gro-(2 comes from 1 alpha Gal)-P]n, in which digalactosyl moieties connect the alpha-galactosylated glycerophosphate units. Principally, the same results were obtained with the more complex system of autolysis of S. aureus cells. We hypothesize that the anti-autolytic activity of LTA resides in a sequence of glycerophosphate units and that the negative charges of appropriately spaced phosphodiester groups play a crucial role. The alanine ester effect is discussed with respect to the putative in vivo regulation of autolysins by LTA.

Alanine↗

[The effects of "relaxing music" on patients, doctors and nursing staff of a medical intensive care unit (author's transl)].

With the intention of reducing the psychological stress and anxiety of patients during their admission to the medical intensive care unit, specially selected and prepared instrumental music was played over a loud speaker to the patient cubicles. The patients' opinions on the programme and their subjective state of health were determined by a questionnaire. 78% of the patients felt that their well-being was improved by the music. With the exception of 7%, who were disturbed by the music, all other patients found it reassuring, diverting, hypnotic or entertaining. In addition to the patients 11 doctors and 16 nurses from the intensive care unit as well as 38 doctors and 80 nurses from 39 medical intensive care units in German university clinics and large hospitals were asked for their opinions on the use of music for intensive care patients.

Acute Disease↗

Glucosamine itself mediates reversible inhibition of protein glycosylation. A study of glucosamine metabolism at inhibitory concentrations in influenza-virus-infected cells.

The metabolism of glucosamine in chick embryo fibroblasts was studied at different concentrations of the amino sugar added to the culture medium. In glucose-containing medium the well-known metabolites, UDP-N-acetylglucosamine, N-acetylglucosamine 6-phosphate and N-acetylglucosamine, are detectable after inhibition of glycosylation resulting from glucosamine treatment. Especially when the cells were infected with influenza virus, high intracellular concentrations of non-metabolized glucosamine are demonstrable in addition. Removal of the inhibitor from the medium results in release of the block of influenza virus glycoprotein glycosylation within 10 min. The onset of glycosylation is paralleled by a rapid reduction of intracellular levels of glucosamine without significant changes in the concentration of its metabolites. Furthermore, concentrations of GDP-mannose, UDP-glucose, and UDP-galactose remain constant for at least 30 min after reversal of the block. It is concluded that glucosamine as such exerts its effect on glycosylation, rather than one of its metabolites being responsible for this effect.

Animals↗

Acyldiglucosyldiacylglycerol-containing lipoteichoic acid with a poly(3-O-galabiosyl-2-O-galactosyl-sn-glycero-1-phosphate) chain from Streptococcus lactis Kiel 42172.

The lipoteichoic acid of Streptococcus lactis Kiel 42172 was isolated. The lipid portions were released by HF and were established to be 3-O-[O-alpha-D-galactopyranosyl-(1 leads to 6)-alpha-D-galactopyranosyl]-2-O-alpha-D-galactopyranosyl-sn-glycero-1-phosphate, they are joined by phosphodiester bonds nosyl)]glycerol. The repeating units of the hydrophilic chain were established to be 3-O-[O-alpha-D-galactopyranosyl-(1 leads to 6)-alpha-D-galactopyranosyl]-2-O-alpha-D-galactopyranosyl-sn-glycero-1-phosphate; they are joined by phosphodiester bonds at carbon atom 6 of the galabiosyl residues. The innermost unit is linked to the glycolipid by a phosphodiester presumably at C-6 of the outer glucosyl moiety. The hydrophilic chain is 7.4--11.8 units in length, measuring 12--19 nm is extended conformation. The content of 2.7--2.96 acyl groups per 2 glucosyl residues indicates that 70--96% of the glycolipid consists of acyldiglucosyldiacylglycerol. The novel poly(glycosylgly-cerophosphate) structure provided for the first time the oplipoteichoic acids are the sn-1 isomer which has previously been suggested from biosynthetic studies (Glaser, L., & Lindsay, B. (1974) Biochem. Biophys. Res. Commun. 59, 1131--1136).

Carbohydrates↗