期刊
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES
卷 1758, 期 1, 页码 38-44出版社
ELSEVIER
DOI: 10.1016/j.bbamem.2006.01.001
关键词
amphotericin B; phospholipid; apolipoprotein; Spectroscopy; nanodisk
资金
- NATIONAL HEART, LUNG, AND BLOOD INSTITUTE [R01HL064159, R37HL064159] Funding Source: NIH RePORTER
- NHLBI NIH HHS [HL-64159] Funding Source: Medline
- NIAID NIH HHS [AI-062541] Funding Source: Medline
Nanodisks (ND) are discrete nanometer scale phospholipid bilayers whose perimeter is circumscribed by amphipathic apolipoproteins. The membranous environment of ND serves as a matrix for solubilizing the polyene antibiotic amphotericin B (AMB). The spectral properties of AMB in ND are dependent upon AMB concentration. Whereas AMB-ND prepared at a concentration of 2.5 mg AMB per 10 mg phospholipid are consistent with AMB self association in the ND membrane environment, AMB-ND prepared at 0.25 or 0.025 mg AMB per 10 mg phospholipid give rise to spectra reminiscent of AMB in organic solvent. Incubation of ND prepared at a phospholipid/AMB ratio of 400:1 (w/w) at 37 degrees C for 1 h induced a shift in absorbance and near UV circular dichroism spectra consistent with antibiotic self-association. The kinetics of this spectral transition were investigated as a function of incubation temperature. While no change in A(388) nm occurred in incubations at 20 degrees C, a time-dependent decrease in A(388) mn was observed at 25, 30 and 37 degrees C. Inclusion of ergosterol in the ND membrane attenuated temperature-induced AMB spectral changes. In Saccharomyces cerevisiae growth inhibition assays, ND containing self associated AMB were somewhat less effective than ND possessing a greater proportion of monomeric AMB. On the other hand, inclusion of ergosterol or cholesterol in the ND particle did not alter the growth inhibition properties of AMB-ND. The miniature membrane environment of ND provides a novel milieu for solubilization and characterization of lipophilic biomolecules. (c) 2006 Elsevier B.V. All rights reserved.
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