Physiology of Neisseria gonorrhoeae

Morse, Stephen; Cacciapuoti, Anthony; Lysko, Paul G.

doi:10.1016/s0065-2911(08)60209-x

Cited by 27 publications

(28 citation statements)

References 196 publications

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“…Although blebbing appeared somewhat more prominent in the resistant bacteria than in the sensitive strain, the significance is uncertain. Blebbing of membranes has also been reported previously for the gonococcus (32).…”

Section: Discussionsupporting

confidence: 74%

Activation of complement by serum-resistant Neisseria gonorrhoeae. Assembly of the membrane attack complex without subsequent cell death.

Harriman

Podack

Braude

et al. 1982

The Journal of Experimental Medicine

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Interaction of the human complement system in normal human serum (NHS) with serum-resistant and -sensitive Neisseria gonorrhoeae was evaluated to better understand the mechanism of serum-resistance. Complement activity (CH50) was depleted from NHS in a dose-dependent fashion by both serum-resistant and -sensitive N. gonorrhoeae. No detectable CH50 remained in NHS incubated with 10(9) colony-forming units (CFU)/ml serum of either resistant or sensitive strains. When smaller numbers of bacteria were incubated with NHS, lesser, yet comparable, amounts of CH50 were depleted by both resistant and sensitive strains. Hemolytic C2 activity was diminished by 33% in the case of resistant N. gonorrhoeae (10(8) CFU/ml serum) and by 48% in the case of a sensitive strain. No detectable decreases in hemolytic C4 or C7 activities were found with either sensitive or resistant strains at this concentration. Both resistant and sensitive strains activated C1s in NHS. Resistant strains specifically activated 19-21% of radiolabeled C1s in NHS, whereas sensitive strains activated 18-32%. Both resistant and sensitive strains also activated C5 in NHS. In binding assays using radiolabeled C5 and C9 in NHS, resistant and sensitive strains bound comparable amounts of C5 and C9. The number of bound C5 and C9 molecules varied according to the number of bacteria or amount of serum used in the assay. The ratio of C9/C5 bound to a sensitive strain was 6.8, and to a resistant strain was 8.2, suggesting that C5 and C9 were incorporated into membrane attack complexes (MAC). Electron microscopic examination of resistant and sensitive strains incubated with NHS revealed that MAC is bound to the surfaces of the resistant strain as well as the sensitive strain.

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Section: Discussionsupporting

confidence: 74%

Activation of complement by serum-resistant Neisseria gonorrhoeae. Assembly of the membrane attack complex without subsequent cell death.

Harriman

Podack

Braude

et al. 1982

The Journal of Experimental Medicine

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“…The ability of N. gonorrhoeae to utilize lactate as a substrate for electron transport has been appreciated for more than 50 years (36,37). The organism possesses an electron transport linked (NAD-independent) L(+) and D(-) lactate dehydrogenase (LDH) associated with the cytoplasmic membrane (1,38,39) as well as a cytosolic NAD-dependent LDH (1). Since lactate derived from mammalian sources is L(+) (40), a gonococcal L(+) LDH is most important in the metabolic responses observed.…”

Section: Resultsmentioning

confidence: 99%

“…Survival demands that the organism acquire substrates critical for metabolism and growth while avoiding or combatting local host defenses. Although much is known about the metabolism and physiology of N. gonorrhoeae (1), the relationship between these characteristics and pathogenesis has received remarkably little attention. Most studies have focused on the role of gonococcal outer membrane components in this process (reviewed in reference 2).…”

Section: Introductionmentioning

confidence: 99%

“…Optimal microbicidal activity of human neutrophils is dependent on their reduction of ambient 02 to superoxide and other reactive oxygen intermediates (13). 02 also plays an integral role in the metabolism of N. gonorrhoeae, being utilized as a terminal electron acceptor under both aerobic and microaerophilic conditions (1,14). Under some experimental conditions, gonococci compete effectively with neutrophils for ambient 02, resulting in diminution of neutrophil formation ofreactive oxygen intermediates (15).…”

Section: Introductionmentioning

confidence: 99%

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Phagocyte-derived lactate stimulates oxygen consumption by Neisseria gonorrhoeae. An unrecognized aspect of the oxygen metabolism of phagocytosis.

Britigan¹,

Klapper²,

Svendsen³

et al. 1988

J. Clin. Invest.

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02 consumption resulting from interaction of Neisseria gonorrhoeae and human neutrophils represents a composite of 02 consumed by the two cell systems. Experiments studying the relative contribution of each system suggested the possibility that gonococci increased their metabolic activity in response to interaction with neutrophils. This hypothesis was confirmed by demonstrating that undifferentiated HL-60 cells, which are unable to undergo a respiratory burst, induce a two-to threefold increase in gonococcal 02 consumption. Gonococcal capacity to adhere to HL-60 cells did not correlate with extent of metabolic stimulation. Stimulatory activity was demonstrable in cell-free supernatant from neutrophils or HL-60 cells, and increased with duration of incubation. Supernatant applied to a G-15 Sephadex column yielded fractions that stimulated gonococcal 02 consumption. Elution profiles were similar for HL-60 cells, neutrophils, and a stimulatory factor previously isolated from pooled human serum. This stimulatory factor(s) failed to adhere to DEAE or C-18 HPLC columns. Stimulatory activity release from myeloid cells was inhibited by incubation at 4°C or in the presence of NaF, indicating a critical role for glucose metabolism. Lactate, the principal product of resting neutrophil glucose catabolism, was demonstrable in cell-free supernatants after incubation at 37°C. Lactate accumulation was inhibited by NaF and decreased temperature of incubation. Lactate at levels present in cell-free supernatant increased gonococcal 02 consumption twofold and restored stimulatory activity to dialyzed serum. Live, but not heat-killed gonococci eliminated lactate released from neutrophils during phagocytosis. Gonococci are able to utilize host-derived lactate to enhance their rate of 02 metabolism.

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“…A nonpiliated, opaque colonial variant of N. gonorrhoeae 1384 (22,23) was used for all studies. Growth conditions were as previously described (23) except that 5.5 mM glucose was the only carbon source used in these studies.…”

mentioning

confidence: 99%

A dominant sulfhydryl-containing protein in the outer membrane of Neisseria gonorrhoeae

et al. 1993

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By using a method that labels sulfhydryl-containing proteins in situ, we (6,14,15). It reacts selectively with the thiol anion of sulfhydryls, crosses biological membranes, and does not significantly alter the molecular weight of proteins (6,14,15).We have used mB to label the sulfhydryl-containing proteins of the outer membrane of Neissena gonorrhoeae. By labeling during growth, we have eliminated the effect of oxidative changes that occur during harvesting and preparation of fractions, thereby detecting the sulfhydryl status of proteins in situ.A nonpiliated, opaque colonial variant of N. gonorrhoeae 1384 (22, 23) was used for all studies. Growth conditions were as previously described (23) (Pharmacia, Piscataway, N.J.) with 0.9 mM dithiothreitol for 30 min and 5.4 mM mB for 20 min. The treated standards were diluted into sample buffer with mercaptoethanol (ME).Electrophoretic analysis of outer membranes from labeled samples revealed multiple fluorescent bands, with the two major fluorescent bands having apparent molecular masses of 41 and 31.5 kDa (Fig. 1A and B). On the basis of its intense fluorescence, the protein at 41 kDa is the major sulfhydryl-containing protein in the outer membrane. Gels stained for protein also showed the 41-kDa protein to be a major protein band in labeled fractions (Fig. 1D, lanes 3 and 5), with an intensity equivalent to that of PIII as seen in outer membrane fractions prepared by conventional methods. However, unlabeled fractions that were not reduced by ME showed no evidence of this protein (Fig. 1D, lanes 1 and 4). When unlabeled samples were reduced, only a lightly staining protein band at 41 kDa was seen (Fig. 1C, lanes 1 and 4). Samples obtained from different experiments (Fig. 1, lanes 1 to 3 versus lanes 4 and 5) confirmed the differences in the 41-kDa protein between labeled and unlabeled samples. Although there was some variation in the intensity of the 41-kDa protein between the two unlabeled, reduced samples, in both cases there was significantly less protein than in labeled samples.There are several possible reasons for the difference between the labeled and unlabeled samples. A likely explanation, given the nature of sulfhydryl compounds, the nature of the reaction with mB, and the effect of ME on the 41-kDa protein in the unlabeled samples, is that in the unlabeled samples artifacts of distribution occur during preparation of the fractions and result in decreases of the 41-kDa protein in the outer membrane; i.e., the protein undergoes autoxidation, with a change in conformation and loss from the membrane, whereas in labeled samples the 41-kDa protein is stable because of its reaction with mB. Another possibility is

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Physiology of Neisseria gonorrhoeae

Cited by 27 publications

References 196 publications

Activation of complement by serum-resistant Neisseria gonorrhoeae. Assembly of the membrane attack complex without subsequent cell death.

Activation of complement by serum-resistant Neisseria gonorrhoeae. Assembly of the membrane attack complex without subsequent cell death.

Phagocyte-derived lactate stimulates oxygen consumption by Neisseria gonorrhoeae. An unrecognized aspect of the oxygen metabolism of phagocytosis.

A dominant sulfhydryl-containing protein in the outer membrane of Neisseria gonorrhoeae

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