During screening of protein-protein interactions, using human protein arrays carrying 19,676 recombinant glutathione s-transferase (GST)-fused human proteins, we identified the high-mobility protein group 20A (HMG20A) as a novel S100A6 binding partner. We confirmed the Ca2+-dependent interaction of HMG20A with S100A6 by the protein array method, biotinylated S100A6 overlay, and GST-pulldown assay in vitro and in transfected COS-7 cells. Co-immunoprecipitation of S100A6 with HMG20A from HeLa cells in a Ca2+-dependent manner revealed the physiological relevance of the S100A6/HMG20A interaction. In addition, HMG20A has the ability to interact with S100A1, S100A2, and S100B in a Ca2+-dependent manner, but not with S100A4, A11, A12, and calmodulin. S100A6 binding experiments using various HMG20A mutants revealed that Ca2+/S100A6 interacts with the C-terminal region (residues 311–342) of HMG20A with stoichiometric binding (HMG20A:S100A6 dimer = 1:1). This was confirmed by the fact that a GST-HMG20A mutant lacking the S100A6 binding region (residues 311–347, HMG20A-ΔC) failed to interact with endogenous S100A6 in transfected COS-7 cells, unlike wild-type HMG20A. Taken together, these results identify, for the first time, HMG20A as a target of Ca2+/S100 proteins, and may suggest a novel linkage between Ca2+/S100 protein signaling and HMG20A function, including in the regulation of neural differentiation.
Furan compounds, especially 5-hydroxymethylfurfural (5-HMF) and furfural (FF), are promising platform chemicals that can be produced from various lignocellulosics. This study presents production of the furan compounds from moso bamboo (Phyllostachys edulis) in an ionic liquid, 1-methylimidazolium hydrogen sulfate ([HMIM]HSO4), which acts as both solvent and acid catalyst during the furan compound production. Vacuum steam distillation (VSD) technique, which facilitates simultaneous removal and purification of the furan compounds, was combined to the process. Reaction of the bamboo sample (30 mg) in [HMIM]HSO4 (3.0 g) at 180 o C for 30 min at 20 hPa with continuous steam supply (3.1 mL/min based on liquid water amount) produced 5-HMF and FF in 20.6 and 11.4 wt% yields, which were significantly greater than the maximum yields of those compounds obtained in the experiment without the VSD equipment (5-HMF: 6.6 wt%, FF: 4.6 wt%). The furan compounds were recovered in a cold trap separated from the main reactor as an aqueous solution with significant purity. After 9 times recycling of the ionic liquid without any refreshing treatment of the medium, the yield of the furan compounds decreased by only 10~30 % compared to the initial yield with fresh [HMIM]HSO4. The small decrease in the yield was attributed mainly to the presence of bamboo-derived solid residue piled in [HMIM]HSO4 during the reaction.
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