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The effect of neomycin and streptomycin on the electrical polarisability of aqueous suspensions of Escherichia coli.

Aqueous suspensions of bacteria scatter light strongly. In addition, the bacteria exhibit strong induced dipole moments in an electric field. In this note we report how, by measuring the intensity of the scattered light, the electric polarisability (alpha) of Escherichia coli could be monitored as small quantities of antibiotics were added to the suspensions. The effect of the presence of quite small quantities of antibiotic on the electrical polarisability, which gave rise to the induced dipole, was dramatic. From the hypothesis that alpha had its origins in the bacteria-solvent interface, a theory is presented which adequately accounts for both alpha and its changes in the presence of these antibiotics. The study is taken to suggest that the antibiotic molecules were adsorbed on to the bacterial surface thereby reducing the surface charge. This in turn reduces the number of counterions and the apparent induced dipole moment. Because the electric-field scattering method is both quick and sensitive to changes in alpha, it may prove a valuable method for the study of antibiotic action on cell and microorganism surfaces.

Escherichia coli↗

Characterisation of aminoglycoside acetyltransferase-encoding genes of neomycin-producing Micromonospora chalcea and Streptomyces fradiae.

Two genes (aac) encoding aminoglycoside-N-acetyltransferase from Streptomyces fradiae and Micromonospora chalcea were cloned: the former identified by hybridization with a homologous gene from Streptomyces rimosus forma paromomycinus, the second by direct expression in Streptomyces lividans using pIJ702 as a vector. These two genes showed pronounced nucleotide and amino acid sequence similarities between themselves and also between previously described streptomycetes aac genes. Comparison of the flanking sequence of actinomycetes aac genes indicates considerable divergence, contrary to the notion that clustered biosynthetic genes for structurally related antibiotics were disseminated in their entirety between microbial species.

Acetyltransferases↗

Phosphoinositide hydrolysis by phospholipase C modulated by multivalent cations La(3+), Al(3+), neomycin, polyamines, and melittin.

Second messenger production from phosphoinositide hydrolysis is regulated by different pathways, such as G-proteins or tyrosine phosphorylation of phosphoinositide phospholipase C (PI-PLC). Another means of altering the activity of PI-PLC is through cation interaction with the phosphoinositide substrate. A variety of organic and inorganic multi-valent cations were examined for their effects on the activity of purified PI-PLC delta. Surprisingly, the cations produced both stimulation and inhibition of PI-PLC catalyzed phosphoinositide hydrolysis, depending on the substrate and the ion to phosphoinositide stoichiometry. These data support the hypothesis that ionic complexes with phosphoinositides may alter their hydrolysis by PI-PLC.

Aluminum↗

A strategy for the design of selective RNA binding agents. Preparation and RRE RNA binding affinities of a neomycin-peptide nucleic acid heteroconjugate library.

We have successfully developed a new strategy for RNA ligand design, which applies the antisense concept to enhance and make more specific loop region interactions while at the same time preserving stem region anchoring. The heteroconjugates, prepared in this effort, have proven to be the most specific small molecule ligands against RRE RNA that have been uncovered to date.

Drug Design↗

Isolation and characterization of a neomycin-resistant mutant of Methanobacterium thermoautotrophicum with a lesion in Na+-translocating ATPase (synthase).

A mutant of Methanobacterium thermoautotrophicum with a lesion in membrane Na+-translocating ATPase (synthase) was isolated. The total ATPase activity in permeabilized cells of this mutant was elevated three-fold as compared with the wild-type strain. In contrast to wild-type cells, mutant ATPase was neither inhibited by DCCD nor stimulated by Na+ ions. The methane formation orate of the mutant cells at pH 7.5 under non-growing conditions was nearly twice that of the wild-type strain and was stimulated by sodium ions. On the other hand, the ATP synthesis driven by methanogenesis under the same conditions was lower that of the wild-type under the same conditions, and contrary to the wild-type was not stimulated by Na+ ions. ATP synthesis driven by a potassium diffusion potential in the presence of sodium ions was markedly diminished in the mutant cells. The membrane potential values of the wild-type and the mutant cells in the presence of 10 mM NaCl at pH 7.0 were comparable at energized conditions (-223 mV and -230 mV respectively). The Mg2+-dependent ATPase activity of the 10(5) x g supernatant of broken cells from the mutant cells was 30% higher than in the wild-type. On the other hand, two bands with Mg2+-dependent ATPase activity were identified by native PAGE in this fraction in both wild-type as well as in mutant. These data suggest that the binding of Na+-translocating ATPase (synthase) to the membrane spanning part is changed in the mutant strain.

Adenosine Triphosphatases↗

Determination of the relative amounts of the B and C components of neomycin by ion-exclusion chromatography using refractometric detection.

Refractometric detection can be used as a convenient alternative to ninhydrin colorimetric or polarimetric detection in ion-exclusion chromatography of neomycinn. The determination of the relative amounts of neomyci B and C using different detection methods is examined. The use of a resin of smaller granulometry and a medium-pressure chromatographic apparatus reduces the analysis time to less than 25 min.

Chromatography, Gel↗