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John H Ipsen

Publications and source records attributed to John H Ipsen.

5 recordsLinked to original sources

Effects of farnesol on the physical properties of DMPC membranes.

Farnesol interacts with membranes in a wide variety of biological contexts, yet our understanding of how it affects lipid bilayers is not yet complete. This study investigates how the 15-carbon isoprenoid, farnesol, influences the phase behaviour, lateral organization, and mechanical stability of dimyristol phosphatidylcholine (DMPC) model membranes. Differential scanning calorimetry (DSC) of multilamellar DMPC-farnesol mixtures (up to 26 mol% farnesol) demonstrates how this isoprenoid lowers and broadens the gel-fluid phase transition. A gel-fluid coexistence region becomes progressively more dominant with increasing farnesol concentration and at concentrations of and greater than 10.8 mol%, an upper transition emerges at about 35 degrees C. Atomic force microscopy images of supported farnesol-DMPC bilayers containing 10 and 20 mol% farnesol provide structural evidence of gel-fluid coexistence around the main transition. Above this coexistence region, membranes exhibit homogeneous lateral organization but at temperatures below the main gel-fluid coexistence region, another form of phase coexistence is observed. The solid nature of the gel phase is confirmed using micropipette aspiration. The combined thermodynamic, structural, and mechanical data allow us to construct a phase diagram. Our results show that farnesol preferentially partitions into the fluid phase and induces phase coexistence in membranes below the main transition of the pure lipid.

Calorimetry, Differential Scanning↗

Vesicle fluctuation analysis of the effects of sterols on membrane bending rigidity.

Sterols are regulators of both biological function and structure. The role of cholesterol in promoting the structural and mechanical stability of membranes is widely recognized. Knowledge of how the related sterols, lanosterol and ergosterol, affect membrane mechanical properties is sparse. This paper presents a comprehensive comparison of the effects of cholesterol, lanosterol, and ergosterol upon the bending elastic properties of 1-palmitoyl-2-oleoyl- sn-glycero-3-phosphocholine giant unilamellar vesicles. Measurements are made using vesicle fluctuation analysis, a nonintrusive technique that we have recently improved for determining membrane bending rigidity. Giving a detailed account of the vesicle fluctuation analysis technique, we describe how the gravitational stabilization of the vesicles enhances image contrast, vesicle yield, and the quality of data. Implications of gravity on vesicle behaviour are also discussed. These recent modifications render vesicle fluctuation analysis an efficient and accurate method for determining how cholesterol, lanosterol, and ergosterol increase membrane bending rigidity.

Computer Simulation↗

Farnesylated peptides in model membranes: a biophysical investigation.

Protein prenylation plays an important role in signal transduction, protein-protein interactions, and the localization and association of proteins with membranes. Using three different techniques, this study physically characterizes the interactions between model dimyristoylphosphatidylcholine membranes and a series of farnesylated peptides. Magic angle spinning nuclear Overhauser enhancement spectroscopy and differential scanning calorimetry reveal that both charged [Ac-Asn-Lys-Asn-Cys-(farnesyl)-OMe and Ac-Asn-Lys-Asn-Cys-(farnesyl)-NH(2)] and uncharged [Ac-Cys-(farnesyl)-OMe and farnesol] species partition into dimyristoylphosphatidylcholine bilayers. Calorimetry and vesicle fluctuation analysis of giant unilamellar vesicles show that the charged peptides modestly decrease the main gel-fluid phase transition and markedly increase the bending rigidity of large unilamellar vesicles. Uncharged species, on the other hand, dramatically decrease the main phase transition and modestly decrease the bending rigidity. No difference with carboxyl methylation is detected.

Biophysics↗

From lanosterol to cholesterol: structural evolution and differential effects on lipid bilayers.

Cholesterol is an important molecular component of the plasma membranes of mammalian cells. Its precursor in the sterol biosynthetic pathway, lanosterol, has been argued by Konrad Bloch (Bloch, K. 1965. Science. 150:19-28; 1983. CRC Crit. Rev. Biochem. 14:47-92; 1994. Blonds in Venetian Paintings, the Nine-Banded Armadillo, and Other Essays in Biochemistry. Yale University Press, New Haven, CT.) to also be a precursor in the molecular evolution of cholesterol. We present a comparative study of the effects of cholesterol and lanosterol on molecular conformational order and phase equilibria of lipid-bilayer membranes. By using deuterium NMR spectroscopy on multilamellar lipid-sterol systems in combination with Monte Carlo simulations of microscopic models of lipid-sterol interactions, we demonstrate that the evolution in the molecular chemistry from lanosterol to cholesterol is manifested in the model lipid-sterol membranes by an increase in the ability of the sterols to promote and stabilize a particular membrane phase, the liquid-ordered phase, and to induce collective order in the acyl-chain conformations of lipid molecules. We also discuss the biological relevance of our results, in particular in the context of membrane domains and rafts.

Calorimetry, Differential Scanning↗