Convergent and stereoselective synthetic routes to Man9GlcNAc2 (1b), alpha-Glc1M9GlcNAc2 (2b), and its stereoisomer beta-Glc1M9GlcNAc2 (3) were established. Interaction analysis of 2b with CRT was measured by 1H NMR spectroscopy, and the first NMR-based evidence for the specific binding of CRT to 2b was obtained.
C-mannosylation is a unique type of protein glycosylation via C-C linkage between an a-mannose and a tryptophan residue. This modification has been identified in about 30 proteins and regulates several functions, such as protein secretion and intracellular localization, as well as protein stability. About half of C-mannosylated proteins are categorized as proteins containing thrombospondin type 1 repeat domain or type I cytokine receptors. To evaluate whether C-mannosylation broadly affects protein functions regardless of protein domain or family, we have sought to identify other types of C-mannosylated protein and analyse their functions. In this study, we focused on receptor activity modifying protein 1, which neither contains thrombospondin type 1 repeat domain nor belongs to the type I cytokine receptors. Our mass spectrometry analysis demonstrated that RAMP1 is C-mannosylated at Trp 56 . It has been shown that RAMP1 transports to the plasma membrane after dimerization with calcitonin receptor-like receptor and is important for ligand-dependent downstream signalling activation. Our results showed that C-mannosylation has no effect on this transport activity. On the other hand, C-mannosylation did enhance protein stability and cell migration activity. Our data may provide new insight into both C-mannosylation research and novel RAMP1 analysis.
A series of cylinder drag experiments were conducted using the density-matching Poly-Styrene Beads particle-fluids mixture to study the flow characteristics of liquefied sands. PIV technique was used to visualize the velocity fields around the moving cylinder. SPH (Smoothed Particle Hydrodynamics) simulations were also conducted to compare with the experimental results. Experiments results show that the solid fraction of 0.555 is quite important in this particle fluid mixture because the drag force exerted on the cylinder increases sharply with the solid fraction if it is greater than this value. This value coincides with the random loose packing density of mono-disperse spheres. PIV analysis shows that the velocity fields are quite localized around the cylinder, and the localized zone is expanded more in the moving direction than in the perpendicular direction. On the other hand, it turned out that the SPH simulations with a simple viscous fluid model cannot reproduce the similar velocity field, which indicates this particle-fluid mixture cannot be regarded as viscous fluid.
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