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Gramicidin Lateral Distribution in Phospholipid Membranes: Fluorescence Phasor Plots and Statistical Mechanical Model
摘要： When using small mole fraction increments to study gramicidins in phospholipid membranes, we found that the phasor dots of intrinsic ?uorescence of gramicidin D and gramicidin A in dimyristoyl-sn-glycero-3-phosphocholine (DMPC) unilamellar and multilamellar vesicles exhibit a biphasic change with peptide content at 0.143 gramicidin mole fraction. To understand this phenomenon, we developed a statistical mechanical model of gramicidin/DMPC mixtures. Our model assumes a sludge-like mixture of ?uid phase and aggregates of rigid clusters. In the ?uid phase, gramicidin monomers are randomly distributed. A rigid cluster is formed by a gramicidin dimer and DMPC molecules that are condensed to the dimer, following particular stoichiometries (critical gramicidin mole fractions, Xcr including 0.143). Rigid clusters form aggregates in which gramicidin dimers are regularly distributed, in some cases, even to superlattices. At Xcr, the size of cluster aggregates and regular distributions reach a local maximum. Before a similar model was developed for cholesterol/DMPC mixtures (Sugar and Chong (2012) J. Am. Chem. Soc. 134, 1164–1171) and here the similarities and differences are discussed between these two models.
关键词： lipid bilayers,membrane organization,gramicidins,statistical mechanics,?uorescence spectroscopy,peptide-lipid interactions
更新于2025-09-23 15:21:21
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Chemical cross-linking of a variety of green fluorescent proteins as F?rster resonance energy transfer donors for Yukon orange fluorescent protein: A project-based undergraduate laboratory experience
摘要： F?rster resonance energy transfer (FRET) is the basis for many techniques used in biomedical research. Due to its wide use in molecular sensing, FRET is commonly introduced in many biology, chemistry, and physics courses. While FRET is of great importance in the biophysical sciences, the complexity and dif?culty of constructing FRET experiments has resulted in limited usage in undergraduate laboratory settings. Here, we present a practical undergraduate laboratory experiment for teaching FRET using a diverse set of green-emitting ?uorescent proteins (FPs) as donors for a cross-linked Yukon orange FP. This laboratory experiment enables students to make the connection of basic lab procedures to real world applications and can be applied to molecular biology, biochemistry, physical chemistry, and biophysical laboratory courses.
关键词： Upper-division undergraduate,orange ?uorescent protein,?uorescence lifetimes,proteins,green ?uorescent protein,biochemistry,TCSPC,?uorescence spectroscopy
更新于2025-09-10 09:29:36