Atomic force microscopy has been used to study the distribution of ganglioside GM1 in model membranes composed of ternary lipid mixtures that mimic the composition of lipid rafts. The results demonstrate that addition of 1% GM1 to 1:1:1 sphingomyelin/dioleoylphosphatidylcholine/cholesterol monolayers leads to the formation of small ganglioside-rich microdomains (40-100 nm in size) that are localized preferentially in the more ordered sphingomyelin/cholesterol-rich phase. With 5% GM1 some GM1 microdomains are also detected in the dioleoylphosphatidylcholine-rich phase. A similar preferential localization of GM1 in the ordered phase is observed for bilayers with the same ternary lipid mixture in the upper leaflet. The small GM1-rich domains observed in these experiments are similar to the sizes for lipid rafts in natural membranes but considerably smaller than the ordered bilayer domains that have been shown to be enriched in GM1 in recent fluorescence microscopy studies of lipid bilayers. The combined data from a number of studies of model membranes indicate that lateral organization occurs on a variety of length scales and mimics many of the properties of natural membranes.
Key indicators: single-crystal X-ray study; T = 293 K; mean (P-O) = 0.001 A ˚ ; disorder in solvent or counterion; R factor = 0.032; wR factor = 0.075; data-to-parameter ratio = 36.0. Single crystals of the title compound, trisodium divanadium-(III) tris(orthophosphate), were grown from a self-flux in the system Na 4 P 2 O 7-NaVP 2 O 7. Na 3 V 2 (PO 4) 3 belongs to the family of NASICON-related structures and is built up from isolated [VO 6 ] octahedra (3. symmetry) and [PO 4 ] tetrahedra (.2 symmetry) interlinked via corners to establish the framework anion [V 2 (PO 4) 3 ] 3À. The two independent Na + cations are partially occupied [site-occupancy factors = 0.805 (18) and 0.731 (7)] and are located in channels with two different oxygen environments, viz sixfold coordination for the first (3. symmetry) and eightfold for the second (.2 symmetry) Na + cation. Related literature For structures and properties of complex phosphates with general formula Na 3 M III 2 (PO 4) 3 (M III = Sc, Fe, Cr), see: Collin et al. (1986); Genkina et al. (1991); Lazoryak et al. (1980); Lucazeau et al. (1986); Masquelier et al. (1992); Susman et al. (1983). For preparation of NaVP 2 O 7 which was used as an educt for crystal growth of the title compound, see: Zatovsky et al. (1999). Experimental Crystal data Na 3 V 2 (PO 4) 3 M r = 455.76 Trigonal, R3c a = 8.7288 (2) A ˚ c = 21.8042 (7) A ˚ V = 1438.73 (7) A ˚ 3 Z = 6 Mo K radiation = 2.66 mm À1 T = 293 K 0.20 Â 0.15 Â 0.10 mm Data collection Oxford Diffraction Xcalibur-3 CCD diffractometer Absorption correction: multi-scan (Blessing, 1995) T min = 0.635, T max = 0.780 12580 measured reflections 1331 independent reflections 1153 reflections with I > 2(I) R int = 0.063
The dissociation of ligand and receptor involves multiple transitions between intermediate states formed during the unbinding process. In this paper, we explored the energy landscape of the streptavidin-biotin interaction by using the atomic force microscope (AFM) to measure the unbinding dynamics of individual ligand-receptor complexes. The rupture force of the streptavidin-biotin bond increased more than 2-fold over a range of loading rates between 100 and 5000 pN/s. Moreover, the force measurements showed two regimes of loading in the streptavidin-biotin force spectrum, revealing the presence of two activation barriers in the unbinding process. Parallel experiments carried out with a streptavidin mutant (W120F) were used to investigate the molecular determinants of the activation barriers. From these experiments, we attributed the outer activation barrier in the energy landscape to the molecular interaction of the '3-4' loop of streptavidin that closes behind biotin.
Phosphorothioate nucleotides have emerged as powerful pharmacological substitutes of their native phosphodiester analogs with important translational applications in antisense oligonucleotide (ASO) therapeutics and cyclic dinucleotide (CDN) synthesis. Stereocontrolled installation of this chiral motif has long been hampered by the systemic use of phosphorus(III) [P(III)]-based reagent systems as the sole practical means of oligonucleotide assembly. A fundamentally different approach is described herein: the invention of a P(V)-based reagent platform for programmable, traceless, diastereoselective phosphorus-sulfur incorporation. The power of this reagent system is demonstrated through the robust and stereocontrolled synthesis of various nucleotidic architectures, including ASOs and CDNs, via an efficient, inexpensive, and operationally simple protocol.
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