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S A Morse

Publications and source records attributed to S A Morse.

At least 19 recordsLinked to original sources
PubMed1979

Phospholipid metabolism in Neisseria gonorrhoeae: phospholipid hydrolysis in nongrowing cells.

Hydrolysis of cell envelope phospholipids was demonstrated in cells of both autolytic and nonautolytic strains of Neisseria gonorrhoeae that were labeled during growth in the presence of [3H] acetate. The label incorporated into the cellular phospholipids was located exclusively in the fatty acid acyl side chains. Labeled cells were incubated for 2 hr in N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid buffer, pH 8.5, containing various additions, and then examined for distribution of 3H in lipids. Ca++ selectively stimulated the deacylation of phosphatidylethanolamine (PE), whereas Mn++ stimulated the deacylation of phosphatidylglycerol (PG). Hydrolysis of phosphatidylethanolamine by phospholipase A was accompanied by the accumulation of lysophosphatidylethanolamine (LPE) and free fatty acids in the cells. Free fatty acids accumulated to a greater extent than lysophosphatidylethanolamine, suggesting that the latter was further hydrolyzed to glycerophosphorylethanolamine (GPE) and free fatty acids by a lysophospholipase. Methanol, ethanol, propanol, and isopropanol, added at concentrations which inhibited growth by 50%, stimulated phospholipase A, but not lysophospholipase activity. Differences in heat inactivation, metal ion requirements, and pH optima suggested that phospholipase A activities with phosphatidylethanolamine or phosphatidylglycerol as substrate and lysophospholipase may be separate enzymes.

Buffers↗
PubMed1979

Cell envelope of Neisseria gonorrhoeae CS7: peptidoglycan protein complex.

Treatment of cells grown to exponential phase with 4% sodium dodecyl sulfate for 3 h at 100 degrees C resulted in solubilization of all cellular components except for peptidoglycan. In most strains, cells cultured in liquid gonococcal broth at pH 7.2 yielded a peptidoglycan composed primarily of N-acetylmuramic acid N-acetylglucosamine, alanine, glutamic acid, and diaminopimelic acid in a molar ratio of 1:1:2:1:1. The peptidoglycan in these cells accounted for 1 to 2% (dry weight) of the cells. However, in cells cultured at pH 6.0, the dry weight of peptidoglycan increased to 4 to 13%. Preliminary investigations indicated that the apparent increase in weight is strain dependent and is due in part to associated protein(s). Neisseria gonorrhoeae strain CS7 had elevated amounts of protein associated with the peptidoglycan regardless of growth pH. The peptidoglycan-protein complex could not be dissociated by additional extraction with sodium dodecyl sulfate, 10 M LiCl2, or ethylenediaminetetraacetate or by 7.5% polyacrylamide gel electrophoresis. The complex could be degraded by lysozyme, trypsin, chymotrypsin, Pronase B, and Chalaropsis sp. muramidase.

Amino Acids↗
PubMed1978

Cell envelope of Neisseria gonorrhoeae: phospholipase activity and its relationship to autolysis.

The relationship between conditions which permit or inhibit cell lysis and those which promote phospholipid hydrolysis in Neisseria gonorrhoeae was investigated. Suspension of exponential-phase gonococci in buffer in the absence of divalent cations resulted in autolysis but not in phosphlipid hydrolysis. The addition of Ca2+ or Mg2+ to the buffer inhibited autolysis and markedly stimulated the hydrolysis of phosphatidylethanolamine. Incubation of cells in buffer at pH 6 inhibited both autolysis and phospholipid hydrolysis.

Bacteriolysis↗
PubMed1978

Effect of pH on the growth and glucose metabolism of Neisseria gonorrhoeae.

This study examined the effect of pH on the metabolism of glucose by Neisseria gonorrhoeae. Radiorespirometric studies revealed that cells growing at pH 7.2 or 8.0 metabolized glucose primarily (ca. 80%) via the Entner-Doudoroff pathway. The remainder of the glucose was metabolized via the pentose phosphate pathway (ca. 20%). The tricarboxylic acid cycle was not active during glucose catabolism at either pH 7.2 or 8.0, and acetate accumulated in the medium. Cells growing at pH 6.0 had markedly increased pentose phosphate pathway activity (ca. 50%) and a functioning tricarboxylic acid cycle. The alteration in pathways was not due to differences in growth rate, but to the pH of the medium. Chemical fractionation of labeled cells and total hexose analyses revealed that growth pH markedly affected the composition of the gonococcus.

Acetates↗
PubMed1978

Binding of cholesterol by Neisseria gonorrhoeae.

The binding of [1,2-3H]cholesterol to Neisseria gonorrhoeae CS-7, Pseudomonas aeruginosa, and Salmonella typhimurium (smooth and rough strains) was investigated. The kinetics of cholesterol binding to N. gonorrhoeae CS-7 demonstrated that binding occurred slowly with maximum binding by 10 h. Under optimum conditions, a large percentage (65%) of the added cholesterol was associated with the cells. Chemical fractionation revealed that ca. 98% of the labeled cholesterol was associated with the cell membrane(s). The bound cholesterol was not esterified and was associated primarily with the cytoplasmic membrane. Intact gonococci bound 4 to 30 times more cholesterol than the deep rough mutant S. typhimurium TA1535, the wild-type S. typhimurium DB-21, and P. aeruginosa. In contrast, isolated cell membranes from all organisms rapidly bound cholesterol to the same extent. Therefore, the outer membrane can function as a permeability barrier to cholesterol. Cholesterol binding to both whole cells and isolated cell membranes was influenced by the incubation temperature. The rate of cholesterol binding by whole cells of N. gonorrhoeae decreased markedly at lower temperatures, with almost complete cessation of binding at 0 degrees C. A similar temperature effect on the binding of cholesterol to isolated membranes was not observed. Thus, the effect of temperature on the binding of cholesterol to whole cells was an effect not on the actual binding process but rather on the ability of the cholesterol molecule to penetrate the lipid domain of the gonococcal outer membrane.

Cell Membrane↗
PubMed1978

The biology of the gonococcus.

Gonorrhea has been known since antiquity. Today, this disease is the most commonly reported infectious disease in the U.S. The natural environment of the etiological agent, Neisseria gonorrhoeae, is man. In this host, the organism usually parasitizes mucosal surfaces populated by columnar epithelial cells. Under certain conditions, the gonococcus may disseminate or spread to adjacent organs. The gonococcus is well adapted to its environment and is a successful parasite. Until recently, gonococci were uniformly sensitive to penicilin. However, a plasmid encoding beta-lactamase has been identified in some isolates. Most strains exhibit specific requirements for various amino acids, vitamins, purines, and pyrimidines. Only glucose, pyruvate, and lactate are utilized as sources of energy. Glucose is dissimilated by a combination of the Entner-Doudoroff and pentose phosphate pathways. A tricarboxylic acid cycle is also present and active under certain conditions. Structurally, the cell envelope of the gonococcus resembles that of a typical Gram-negative bacterium. Gonococci are highly autolytic, especially in older cultures or after depletion of the energy source. Autolysis is not due solely to peptidoglycan hydrolysis, but appears to involve a destabilization of the outer membrane as well. Cell surface components such as pili, lipopolysaccharide, outer membrane proteins, and a capsule are associated with the virulence and pathogenicity of this organism.

Amino Acids↗
PubMed1977

Binding of progesterone to Neisseria gonorrhoeae and other gram-negative bacteria.

The binding of [1,2-(3)H]progesterone to progesterone-sensitive Neisseria gonorrhoeae CS-7 and the progesterone-insensitive Neisseria mucosa, Pseudomonas aeruginosa, and Salmonella typhimurium (rough and smooth strains) was investigated. The kinetics of binding to N. gonorrhoeae CS-7 demonstrated that the majority of the progesterone binding occurred and equilibrium was reached within the first 30 min. Despite the rapid binding of progesterone, only about 20% of the added steroid was bound at the cell concentration used throughout this study. Whole cells of progesterone-insensitive bacteria bound progesterone less efficiently than the progesterone-sensitive N. gonorrhoeae CS-7. N. mucosa bound low amounts of this steroid (20% of that bound by N. gonorrhoeae CS-7) whereas the other gram-negative bacteria exhibited little progesterone binding (<3% of that bound by N. gonorrhoeae CS-7). The outer membrane permeability of N. gonorrhoeae CS-7, as measured by crystal violet uptake and inhibition, was similar to the deep rough mutant of S. typhimurium TA 1535. The latter organism neither bound nor was inhibited by progesterone. However, isolated cell envelopes of N. gonorrhoeae and progesterone-insensitive bacteria all bound progesterone equally well. Cortisone and cholesterol, althouh structurally similar to progesterone, were not inhibitory to N. gonorrhoeae and did not bind to whole cells as well as progesterone. The major site of progesterone binding appeared to be the cytoplasmic membrane, which bound four times more progesterone than the outer membrane. In addition, isolated cytoplasmic membrane proteins bound more than three times more progesterone per milligram of protein than the intact membrane.

Cell Membrane↗
PubMed1977

Inhibitory action of fatty acids on the growth of Neisseria gonorrhoeae.

Fatty acids of various chain lengths (C(1) to C(24)) were examined for their effects on growth, oxygen consumption, and in vitro reduced nicotinamide adenine dinucleotide oxidase activity of Neisseria gonorrhoeae CS-7. The growth inhibition caused by saturated fatty acids increased with increasing chain length to a maximum with palmitic acid (C(16)). Stearic acid (C(18)) and longer saturated fatty acids showed little inhibition of growth. However, unsaturated fatty acids of chain length C(16) to C(20) were inhibitory. Similar inhibition was observed with Bacillus subtilis and a deep rough mutant of Salmonella typhimurium. Wildtype S. typhimurium and Pseudomonas aeruginosa were more resistant to medium-chain (C(7) to C(10)) fatty acids and completely resistant to long-chain (C(12) to C(18)) fatty acids. Thus, sensitivity of N. gonorrhoeae to long-chain fatty acids appears to be related to the permeability of the outer membrane. Growth inhibition by short-chain (C(1) to C(6)) fatty acids was pH dependent; inhibition of growth increased with decreasing pH. Saturated fatty acids inhibited oxygen consumption by log-phase cells of N. gonorrhoeae. This inhibition increased with increasing chain length to a maximum observed with myristic acid (C(14)). Whereas stearic acid (C(18)) had little effect upon oxygen consumption, unsaturated C(18) fatty acids were inhibitory. An in vitro inhibition of reduced nicotinamide adenine dinucleotide oxidase activity by saturated (C(1) to C(12)) and unsaturated (C(16) to C(20)) fatty acids was also observed. Although the inhibitory concentrations were generally higher than those required to inhibit growth or oxygen consumption, an inhibition of electron transport may be partially responsible for the observed growth inhibition.

Bacteria↗
PubMed1977

Cell envelope of Neisseria gonorrhoeae: relationship between autolysis in buffer and the hydrolysis of peptidoglycan.

Neisseria gonorrhoeae readily underwent autolysis when suspended in N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES) buffer at alkaline pH values. Autolysis was inhibited by the addition of Mg2+ or other divalent cations. Autolysis was also suppressed at acid pH (pH 6.0). Suspension of cells in buffer was accompanied by the hydrolysis of peptidoglycan. The rate of peptidoglycan hydrolysis in HEPES buffer was maximal at pH 8.5 and was similar in the presence or absence of Mg2+. Therefore, divalent cation stabilization against autolysis is not mediated by inhibition of peptidoglycan hydrolysis. Peptidoglycan hydrolysis occurred in HEPES buffer (pH 6.0), but at a rate that was 50% of the maximum. Incubation of cells with chloramphenicol or rifampin before suspension in HEPES buffer (pH 8.5) partially prevented autolysis; under these conditions, peptidoglycan hydrolysis still occurred, but at a reduced rate. Old and new peptidoglycans were hydrolyzed at similar rates. Peptidoglycan hydrolysis results in solubilization of both the peptide and glycan moieties.

Buffers↗
PubMed1977

Cell envelope of Neisseria gonorrhoeae: penicillin enhancement of peptidoglycan hydrolysis.

The addition of 10 microgram of penicillin G per ml to log-phase cultures of Neisseria gonorrhoeae JW-31 (minimum inhibitory concentration for penicillin G, less than 0.007 microgram/ml) resulted in cellular lysis after a lag of 30 min. Penicillin markedly decreased the rate of peptidoglycan synthesis and enhanced the rate of hydrolysis of existing peptidoglycan. Hydrolysis was initiated immediately after addition of penicillin; cellular lysis did not occur until a considerable percentage of the peptidoglycan had been degraded. Cellular lysis was not due to penicillin per se but resulted from inhibition of cell wall synthesis. When cells were grown in media buffered with N-2-hydroxyethyl piperazine-N'-2-ethanesulfonic acid at pH 6, penicillin did not cause lysis; however, at this pH, peptidoglycan hydrolysis occurred and cells lost viability at the same rate as in the control (pH 7.2). We suggest that the stability of gonococci grown at pH 6 is related to increased stability of the outer membrane. The penicillin-enhanced rate of peptidoglycan hydrolysis decreased approximately 50% at pH 6.0. Penicillin-enhanced lysis, peptidoglycan hydrolysis, and loss of viability were also markedly reduced in cells grown at 28 degrees C.

Bacterial Proteins↗
PubMed1976

Inhibition of Neisseria gonorrhoeae by a bacteriocin from Pseudomonas aeruginosa.

Supernatants from broth-grown cultures of Pseudomonas aeruginosa PA 103 exhibited bactericidal activity against Neisseria gonorrhoeae. The concentration of the bactericidal substance increased significantly after induction by mitomycin C. Purification was effected by salt fractionation, chromatography on diethylaminoethyl-cellulose, and sedimentation by centrifugation at 100,000 x g for 90 min. Electron microscopy of this purified preparation revealed structures resembling R-type pyocins in both the contracted and uncontracted state. Pyocins in the contracted state were observed in association with the gonococcal cell surface. No loss of bactericidal activity was observed after treatment with proteolytic enzymes. Standard pyocin typing procedures identified the pyocin pattern as 611 131. The bactericidal activity of this pyocin was examined on various species of Neisseria. Out of 56 strains of N. gonorrhoeae from disseminated and nondisseminated infections, all were susceptible to pyocin 611 131. However, only 3 of 20 strains of N. meningitidis and 5 of 16 strains of N. lactamica were susceptible. The bactericidal activity that pyocin 611 131 has for N. gonorrhoeae and other species of Neisseria is significant because it departs from the expected specificity that heretofore has distinguished bacteriocins from most "classical" antibiotics.

Bacteriocins↗
PubMed1976

Physiology and metabolism of pathogenic neisseria: tricarboxylic acid cycle activity in Neisseria gonorrhoeae.

Tricarboyxlic acid cycle activity was examined in Neisseria gonorrhoeae CS-7. The catabolism of glucose in N. gonorrheae by a combination of the Entner-Doudoroff and pentose phosphate pathways resulted in the accumulation of acetate, which was not further catabolized until the glucose was depleted or growth became limiting. Radiorespirometric studies revealed that the label in the 1 position of acetate was converted to CO2 at twice the rate of the label in the 2 position, indicating the presence of a tricarboxylic acid cycle. Growth on glucose markedly reduced the levels of all tricarboxylic acid cycle enzymes except citrate synthase (EC 4.1.3.7). Extracts of glucose-grown cells contained detectable levels of all tricarboxylic acid cycle enzymes except aconitase (EC 4.2.1.3), isocitrate dehydrogenase (EC 1.1.1.42), and a pyridine nucleotide-dependent malate dehydrogenase (EC 1.1.1.37). Extracts of cells capable of oxidizing acetate lacked only the pyridine nucleotide-dependent malate dehydrogenase. In lieu of this enzyem, a particulate pyridine nucleotide-independent malate oxidase (EC 1.1.3.3) was present. This enzyme required flavin adenine dinucleotide for activity and appeared to be associated with the electron transport chain. Radiorespirometric studies utilizing labeled glutamate demonstrated that a portion of the tricarboxylic acid cycle functioned during glucose catabolism. In spite of the presence of all tricarboxylic acid cycle enzymes, N. gonorrhoeae CS-7 was unable to grow in medium supplemented with cycle intermediates.

Acetates↗
PubMed1976

Adaptation of the Minitek system for the rapid identification of Neisseria gonorrhoeae.

A suspending medium was developed for use with the Minitek system for the confirmatory identification of Neisseria gonorrhoeae, N. meningitidis, and N. lactamica based upon the production of acid from various carbohydrates. The addition of sodium bicarbonate to the medium made negative reactions easier to read. More isolates of N. gonorrhoeae were identified with the suspending medium in the Minitek system than with cystine-Trypticase agar media. With a suitable inoculum size, a positive identification could be made in less than 1 h; most isolates (90,8%) could be identified within 4 h of inoculation. The Minitek system is reliable and easy to use.

Acids↗