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41.
Voltage-dependent depolarization of bacterial membranes and artificial lipid bilayers by the peptide antibiotic nisin 总被引:24,自引:0,他引:24
The peptide antibiotic nisin is shown to disrupt valinomycin-induced potassium diffusion potentials imposed on intact cells of Staphylococcus cohnii 22. Membrane depolarization occurred rapidly at high diffusion potentials while at low potentials nisin-induced depolarization was slower suggesting that nisin requires a membrane potential for activity. This assumption was proven in experiments with planar lipid bilayers (black lipid membranes). Macroscopic conductivity measurements indicated a voltage-dependent action of nisin. The potential must have a trans-negative orientation with respect to the addition of nisin (added to the cis-side) and a sufficient magnitude (ca. -100 mV). With intact cells the threshold potential was lower (-50 to -80 mV at pH 7.5 and below -50 mV at pH 5.5). Single channel recordings resolved transient multistate pores, strongly resembling those introduced by melittin into artificial bilayers. The pores had diameters in the range of 0.2–1 nm, and lifetimes of few to several hundred milliseconds. The results indicate that nisin has to be regarded as a membrane-depolarizing agent which acts in a voltage-dependent fashion.Abbreviations BLM
Black lipid membranes
- CCCP
carbonyl cyanide m-chlorophenylhydrazone
- DOPC
dioleoyl phosphatidylcholine
- PS
phosphatidylserine
- TPP+
tetraphenylphosphonium cation 相似文献
42.
J. Ruiz-Herrera J. P. Martinez M. Casanova M. L. Gil R. Sentandreu 《Archives of microbiology》1987,149(2):156-162
Cells from the slime variant of Neurospora crassa were broken in isotonic conditions by use of triethanolamine buffer plus EDTA. After removal of large membranous structures by low-speed centrifugation, chitosomes and secretory vesicles were separated by means of gel filtration, precipitation of membranous contaminants with Concanavalin A, and centrifugation in sucrose or glycerol gradients. Polypeptidic composition of fractions enriched in secretory vesicles or chitosomes was found to be distinct. By these criteria we concluded that chitosomes and secretory vesicles represent different populations of microvesicles. Both microvesicular populations appeared free of endoplasmic reticulum and vacuolar contaminants as demonstrated by determination of appropriate enzymatic markers.Abbreviations ER
Endoplasmic reticulum
- UDP-GlcNAc
uridine-5-diphosphate N-acetyl glucosamine
- GlcNAc
N-acetyl glucosamine
- SDS
sodium dodecyl sulfate
- PMSF
phenyl methyl sulfonyl fluoride
- EDTA
ethylene diamino tetraacetic acid
Investigador Nacional de Mexico. On leave from the Centro de Investigacion y Estudios Avanzados (IPN), and the Universidad de Guanajuato, Gto., Mexico 相似文献
43.
T. de la Rubia J. Gonzalez-Lopez J. Moreno M. V. Martinez-Toledo A. Ramos-Cormenzana 《Archives of microbiology》1987,147(4):354-357
The effect of a low phosphate concentration on intracellular adenine nucleotide content, oxygen consumption and poly--hydroxybutyrate deposition was investigated with N-free and NH
4
+
batch cultures of Azotobacter vinelandii. When the microorganisms were cultured under low-phosphate concentrations the cells contained much larger amounts of poly--hydroxybutyrate, but displayed lower oxygen consumption activities and energy charge values than did control cells. Also, the ratio ATP to ADP was much higher in control cells and the intracellular levels of ATP were lower in low-phosphate cells. 相似文献
44.
Dr. Richard Reynolds Christine Steffen Norbert Herschkowitz 《Neurochemical research》1987,12(10):885-890
Oligodendrocytes were isolated from mixed glial cultures of neonatal mouse forebrain and further grown in serum-free hormone supplemented culture medium. Cell populations were identified by indirect immunofluorescence using a range of specific antibodies, revealing a predominantly immature population of oligodendrocytes, the majority expressing the myelin glycolipids galactocerebroside and sulfatide on their plasma membrane. Astroglial contamination was found to be minimal. Simultaneous autoradiography and immunofluorescence demonstrated the presence of a transport system for the major inhibitory neurotransmitter GABA in the oligodendrocytes. The transport system was found to be energy, sodium and temperature dependent. Kinetic analysis revealed a high affinity system, with aK
m of 6.27 M and aV
max of 0.714 nmol/min/mg protein, which is comparable to that found previously for CNS neurons and astrocytes.Special Issue dedicated to Dr. E. M. Shooter and Dr. S. Varon. 相似文献
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Jay C. D. Hinton Julie M. Sidebotham Lizbeth J. Hyman Michel C. M. Pérombelon George P. C. Salmond 《Molecular & general genetics : MGG》1989,217(1):141-148
Summary The blackleg pathogen Erwinia carotovora subsp. atroseptica (Eca) causes an economically important disease of potatoes. We selected a genetically amenable Eca strain for the genetic analysis of virulence. Tn5 mutagenesis was used to generate nine mutants which exhibited reduced virulence (Rvi-) of strain SCRI1043. Following physiological characterisation, mutants were divided into three classes: (1) auxotrophs; (2) extracellular enzyme mutants; and (3) a growth rate mutant. The isolation of these Rvi- mutants has allowed us to consider some factors that affect Eca virulence. 相似文献