Biomedical subjects

M T Lamy-Freund

Publications and source records attributed to M T Lamy-Freund.

22 records · Page 2Linked to original sources

PubMed1991

Cholesterol modulation of lipid-protein interactions in liver microsomal membrane: a spin label study.

ESR spectra of spin probes were used to monitor lipid-protein interactions in native and cholesterol-enriched microsomal membranes. In both systems composite spectra were obtained, one characteristic of bulk bilayer organization and another due to a motionally restricted population, which was ascribed to lipids in a protein microenvironment. Computer spectral subtractions revealed that cholesterol modulates the order/mobility of both populations in opposite ways, i.e., while the lipid bilayer region gives rise to more anisotropic spectra upon cholesterol enrichment, the spectra of the motionally restricted population become indicative of increased mobility and/or decreased order. These events were evidenced by measurement of both effective order parameters and correlation times. The percentages of the motionally restricted component were invariant in native and cholesterol-enriched microsomes. Variable temperature studies also indicated a lack of variation of the percentages of both spectral components, suggesting that the motionally restricted one was not due to protein aggregation. The results correlate well with the effect of cholesterol enrichment on membrane-bound enzyme kinetics and on the behavior of fluorescent probes [Castuma & Brenner (1986) Biochemistry 25, 4733-4738]. Several hypothesis are put forward to explain the molecular mechanism of the cholesterol-induced spectral changes.

Animals↗

PubMed1991

Characterization and time dependence of amphotericin B: deoxycholate aggregation by quasielastic light scattering.

Quasielastic light scattering measurements of amphotericin B (AB):deoxycholate (DOC) preparations provided information about particle size and aggregation as a function of concentration. The data allowed the time dependence of the aggregation to be followed and indicated that the initial rates of the change in average equivalent hydrodynamic diameter increased with decreasing concentration. The results extend the model proposed by Lamy-Freund and co-workers, which describes AB:DOC systems as consisting of AB:DOC mixed aggregates co-existing with pure DOC micelles. Although the AB:DOC aggregates are unstable at all concentrations studied, the rate of aggregation increases by three orders of magnitude as the concentration is reduced from 20 mM (DOC concentration) to the concentration region of DOC micellization. These results are in agreement with the different distribution of AB and DOC in the body of experimental animals, and may be of relevance for the understanding of the serious toxic effects of AB.

Amphotericin B↗

PubMed1989

Polydispersity of aggregates formed by the polyene antibiotic amphotericin B and deoxycholate. A spin label study.

The amphotericin B-deoxycholate (AB-DOC) system (1:2, mole basis) was studied with regard to its organizational properties making use of spin label ESR spectra. The spectra of a fatty acid spin label intercalated in AB-DOC preparations revealed two components, one strongly (S) and one weakly (W) immobilized. Spectral subtractions indicated that S corresponds to label in mixed AB-DOC aggregates while W is due to label in deoxycholate micelles. This situation, coexistence of different aggregates, is similar to that found in systems consisting of bile salts and phospholipids. The DOC/AB mole ratio in the mixed aggregate is highest when pure DOC micelles are present. Dilution leads to disappearance of the latter and to continuous loss of DOC from AB-DOC accompanied by an increase in size and decrease in solubility of the aggregates, as verified by filtration and centrifugation experiments. The results indicate that AB-DOC systems are polydisperse. Since amphotericin B preparations having different organizational properties display different toxic and therapeutic effect, the study of amphotericin B aggregates should help in understanding these phenomena at a molecular level.

Amphotericin B↗

PubMed1985

Mechanism of inactivation of the polyene antibiotic amphotericin B. Evidence for radical formation in the process of autooxidation.

Radical formation during autooxidation of the polyene antibiotic amphotericin B was monitored by following the kinetics of decay of the ESR signal of a stable nitroxide. The kinetics were seen to depend both on antibiotic and on nitroxide concentration. The radicals formed were studied by spin trapping. Three preparations--clinical Fungizone, amphotericin B suspended in buffer, and amphotericin B in buffer - 10% dimethyl sulfoxide--yielded spin adducts of different nature and/or concentrations. In the absence of dimethyl sulfoxide, amphotericin B yielded at least two carbon-centered radicals whose spectra indicated restricted mobility. This suggests that the radicals arise from the whole amphotericin B molecule located in molecular aggregates present in the preparations, and not from smaller and possibly more water-soluble fragments of the antibiotic. In the presence of dimethyl sulfoxide the spin adducts derived from secondary carbon radicals originated from this solvent. Their spectra were indicative of fast tumbling. Direct evidence for autooxidation was obtained by measuring oxygen consumption. All processes examined occurred at the same time scale observed for drug inactivation, in agreement with the idea that loss of activity is due to antibiotic autooxidation.

Amphotericin B↗