separation-spinach-thylakoid-protein-complexes-native-green-gel

Separation of Spinach Thylakoid Protein Complexes by Native Green Gel Electrophoresis and Band Characterization using Time-Correlated Single Photon Counting

The light reactions of photosynthesis are carried out by a series of pigmented protein complexes in the thylakoid membranes. The stoichiometry and ...

Michigan State University

We report on the use of diffusion measurements to gauge the fluidity and surface binding properties of a molecular monolayer. The monolayer film consists of octadecyl-1-phosphonic acid (ODPA) and controlled amounts of a lyso-phosphatidic acid tagged with the fluorescent probe BODIPY (BLPA). The monolayer films were formed using a Langmuir-Blodgett (LB) trough and deposited onto a glass slide. Monolayer morphology was characterized during film formation using Brewster angle microscopy (BAM). Fluorescence Recovery After Photobleaching (FRAP) microscopy was used to measure translational diffusion of BLPA and Fluorescence Anisotropy Decay Imaging (FADI) was used to measure rotational diffusion of the BLPA chromophore. These results provide information on the motional freedom of the probe and, importantly, on the strength of interaction between the probe and the support. Compositional variations in the monolayer give rise to changes in constituent dynamics that reflect intermolecular interactions. 

Diffusional motion as a gauge of fluidity and interfacial adhesion. Supported alkylphosphonate monolayers.

We report on the morphology and dynamics of planar supported lipid bilayer structures as a function of pH and ionic strength of the aqueous overlayer. Supported lipid bilayers composed of three components (phosphocholine, sphingomyelin and cholesterol) are known to exhibit phase segregation, with the characteristic domain sizes dependent on the amount and identity of each constituent, and the composition of the aqueous overlayer in contact with the bilayer. We report on fluorescence anisotropy decay imaging measurements of a rhodamine chromophore tethered to the headgroup of a phosphoethanolamine, where anisotropy decay images were acquired as a function of solution overlayer pH and ionic strength. The data reveal a two-component anisotropy decay under all conditions, with the faster time constant being largely independent of pH and ionic strength and the slower component depending on pH and ionic strength in different manners. For liposomes of the same composition, a single exponential anisotropy decay was seen. We interpret this difference in terms of bilayer curvature and support surface-bilayer interactions, and the pH and ionic strength dependencies in terms of ionic screening and protonation in the bilayer headgroup region. 

Gary Blanchard

Department of Chemistry

Michigan State University

Separation of Spinach Thylakoid Protein Complexes by Native Green Gel Electrophoresis and Band Characterization using Time-Correlated Single Photon Counting

Gary Blanchard

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Chemistry

Michigan State University

Separation of Spinach Thylakoid Protein Complexes by Native Green Gel Electrophoresis and Band Characterization using Time-Correlated Single Photon Counting

Department of Chemistry