The organization of biomolecules and bioassemblies is highly governed by the nature and extent of their interactions with water. These interactions are of high intricacy and a broad range of methods based on various principles have been introduced to characterize them. As these methods view the hydration phenomena differently (e.g., in terms of time and length scales), a detailed insight in each particular technique is to promote the overall understanding of the stunning "hydration world." In this prospective mini-review we therefore critically examine time-dependent fluorescence shift (TDFS)-an experimental method with a high potential for studying the hydration in the biological systems. We demonstrate that TDFS is very useful especially for phospholipid bilayers for mapping the interfacial region formed by the hydrated lipid headgroups. TDFS, when properly applied, reports on the degree of hydration and mobility of the hydrated phospholipid segments in the close vicinity of the fluorophore embedded in the bilayer. Here, the interpretation of the recorded TDFS parameters are thoroughly discussed, also in the context of the findings obtained by other experimental techniques addressing the hydration phenomena (e.g., molecular dynamics simulations, NMR spectroscopy, scattering techniques, etc.). The differences in the interpretations of TDFS outputs between phospholipid biomembranes and proteins are also addressed. Additionally, prerequisites for the successful TDFS application are presented (i.e., the proper choice of fluorescence dye for TDFS studies, and TDFS instrumentation). Finally, the effects of ions and oxidized phospholipids on the bilayer organization and headgroup packing viewed from TDFS perspective are presented as application examples.

• Klíčová slova
• biomembranes, calcium, cholesterol, hydration, lipid headgroups, membrane dynamics, oxidized phosholipids, time-dependent fluorescence shift,
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Understanding interactions of calcium with lipid membranes at the molecular level is of great importance in light of their involvement in calcium signaling, association of proteins with cellular membranes, and membrane fusion. We quantify these interactions in detail by employing a combination of spectroscopic methods with atomistic molecular dynamics simulations. Namely, time-resolved fluorescent spectroscopy of lipid vesicles and vibrational sum frequency spectroscopy of lipid monolayers are used to characterize local binding sites of calcium in zwitterionic and anionic model lipid assemblies, while dynamic light scattering and zeta potential measurements are employed for macroscopic characterization of lipid vesicles in calcium-containing environments. To gain additional atomic-level information, the experiments are complemented by molecular simulations that utilize an accurate force field for calcium ions with scaled charges effectively accounting for electronic polarization effects. We demonstrate that lipid membranes have substantial calcium-binding capacity, with several types of binding sites present. Significantly, the binding mode depends on calcium concentration with important implications for calcium buffering, synaptic plasticity, and protein-membrane association.

• MeSH
• buněčná membrána metabolismus   MeSH
• fosfolipidy chemie   metabolismus   MeSH
• lipidové dvojvrstvy chemie   metabolismus   MeSH
• liposomy chemie   metabolismus   MeSH
• molekulární modely MeSH
• simulace molekulární dynamiky MeSH
• vápník metabolismus   MeSH
• vápníková signalizace MeSH
• vazebná místa MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH
• Názvy látek
• fosfolipidy MeSH
• lipidové dvojvrstvy MeSH
• liposomy MeSH
• vápník MeSH

The need for detailed biophysical description of cationic lipid membranes, which are commonly used as gene transfection vectors, led us to study the properties of mixed cationic/zwitterionic lipid bilayers. Fluorescence solvent relaxation measurements of 6-dodecanoyl-2-dimethylaminonaphthalene (Laurdan) incorporated in a membrane consisting of cationic dimyristoyltrimethylammoniumpropane (DMTAP) and zwitterionic dimyristoylphosphatidylcholine (DMPC) were performed. The obtained results are compared with a recently measured system consisting of dioleoyltrimethylammoniumpropane (DOTAP) and dioleoylphosphatidylcholine (DOPC) (Jurkiewicz et al. Langmuir 22:8741-8749, 2006). The similar nonmonotonic dependence of the relaxation kinetics on cationic lipid content in the membrane was present for both systems. While the slowest solvent relaxation have been observed for 30 mol% of DOTAP in the DOPC bilayer (Jurkiewicz et al. Langmuir 22:8741-8749, 2006), for DMPC/DMTAP system it was found at 45 mol% of DMTAP, which agrees with the literature. Both membranes increased their hydration upon increased cationic lipid content.

• MeSH
• 2-naftylamin analogy a deriváty   chemie   MeSH
• dimyristoylfosfatidylcholin chemie   MeSH
• fluorescence MeSH
• fluorescenční barviva chemie   MeSH
• kationty chemie   MeSH
• kvartérní amoniové sloučeniny chemie   MeSH
• laurany chemie   MeSH
• lipidové dvojvrstvy chemie   MeSH
• lipidy chemie   MeSH
• membrány chemie   MeSH
• molekulární struktura MeSH
• myristáty chemie   MeSH
• rozpouštědla chemie   MeSH
• voda chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 1,2-dimyristoyl-3-trimethylammonium propane MeSH Prohlížeč
• 2-naftylamin MeSH
• dimyristoylfosfatidylcholin MeSH
• fluorescenční barviva MeSH
• kationty MeSH
• kvartérní amoniové sloučeniny MeSH
• laurany MeSH
• laurdan MeSH Prohlížeč
• lipidové dvojvrstvy MeSH
• lipidy MeSH
• myristáty MeSH
• rozpouštědla MeSH
• voda MeSH