Construction of a High‐Mannose‐Type Glycan Library by a Renewed Top‐Down Chemo‐Enzymatic Approach

A comprehensive method for the construction of a high‐mannose‐type glycan library by systematic chemo‐enzymatic trimming of a single Man9‐based precursor was developed. It consists of the chemical synthesis of a non‐natural tridecasaccharide precursor, the orthogonal demasking of the non‐reducing ends, and trimming by glycosidases, which enabled a comprehensive synthesis of high‐mannose‐type glycans in their mono‐ or non‐glucosylated forms. It employed glucose, isopropylidene, and N‐acetylglucosamine groups for blocking the A‐, B‐, and C‐arms, respectively. After systematic trimming of the precursor, thirty‐seven high‐mannose‐type glycans were obtained. The power of the methodology was demonstrated by the enzymatic activity of human recombinant N‐acetylglucosaminyltransferase‐I toward M7–M3 glycans, clarifying the substrate specificity in the context of high‐mannose‐type glycans.     

Approaches toward High‐Mannose‐Type Glycan Libraries

Asparagine‐linked (N‐linked) sugar chains are widely found in the rough endoplasmic reticulum (ER), which has attracted renewed attention because of its participation in the glycoprotein quality control process. In the ER, newly formed glycoproteins are properly folded to higher‐order structures by the action of a variety of lectin chaperones and processing enzymes and are transported into the Golgi, while terminally misfolded glycoproteins are carried into the cytosol for degradation. A group of proteins related to this system are known to recognize subtle differences in the high‐mannose‐type oligosaccharide structures of glycoproteins; however, their molecular foundations are still unclear. In order to gain a more precise understanding, our group has established a strategy for the systematic synthesis of high‐mannose‐type glycans. More recently, we have developed “top‐down” chemoenzymatic approaches that allow expeditious access to theoretically all types of high‐mannose glycans. This strategy comprehensively delivered 37 high‐mannose‐type glycans, including G1M9–M3 glycans, and opened up the possibility of the elucidation of structure–function relationships with a series of high‐mannose‐type glycans.     

β‐Cyclodextrin Pseudopolyrotaxanes with π‐Conjugated Polymer Axles

Summary: β‐Cyclodextrin (β‐CD) pseudopolyrotaxanes containing poly(thiophene‐2,5‐diyl), PTh , or poly(3‐methylthiophene‐2,5‐diyl)s, P3MeTh s, as an axle were prepared. Structures of the pseudopolyrotaxanes and their inclusion behavior with β‐CD were investigated. The UV‐vis measurements revealed that inclusion of P3MeTh s by β‐CD depended on the flexibility of the main chain and their molecular weight.

Formation of the inclusion complex of β‐CD and PTh .     

Evaluation of gadoxetate disodium as a contrast agent for mouse liver imaging: comparison with gadobenate dimeglumine

We investigated the characteristics of gadoxetate disodium (Gd-EOB-DTPA) as a contrast agent for magnetic resonance imaging of the mouse liver. Mice were imaged sequentially under isoflurane anesthesia using a T1-weighted, three-dimensional fast low-angle shot (3D FLASH) sequence after an intravenous injection of Gd-EOB-DTPA or gadobenate dimeglumine (Gd-BOPTA), and the time course of the contrast effect was examined. The time course of the contrast effect of Gd-EOB-DTPA was also assessed after intravenous injection under pentobarbital anesthesia and after subcutaneous injection while awake or under isoflurane or pentobarbital anesthesia. Moreover, different doses of Gd-EOB-DTPA or Gd-BOPTA were injected subcutaneously into conscious mice, and the clarity of the liver border was evaluated visually. Intravenous injection under isoflurane anesthesia caused rapid contrast enhancement in the liver with both Gd-EOB-DTPA and Gd-BOPTA, and the contrast effect was 41% stronger with Gd-EOB-DTPA. Subcutaneous injection of Gd-EOB-DTPA caused delayed but favorable contrast enhancement in the liver. Washout of Gd-EOB-DTPA was faster in mice injected while awake than in those injected under anesthesia. After intravenous injection, washout was faster under pentobarbital anesthesia than under isoflurane anesthesia. The peak liver contrast was 11% and 18% stronger under pentobarbital anesthesia than under isoflurane anesthesia, after intravenous and subcutaneous injections, respectively. Subcutaneous injection of Gd-EOB-DTPA or Gd-BOPTA caused dose-dependent contrast effects in the liver. At a given dose, the contrast effect tended to be stronger and liver demarcation tended to be clearer with Gd-EOB-DTPA than with Gd-BOPTA. In conclusion, intravenous or subcutaneous injection of Gd-EOB-DTPA produces a favorable contrast effects in the mouse liver, indicating its potential in investigating mouse models of liver diseases. The contrast effects vary between conscious mice and anesthetized mice and among anesthetic agents used.     

HPLC-assisted automated oligosaccharide synthesis

A standard HPLC was adapted to polymer supported oligosaccharide synthesis. Solution-based reagents are delivered using a software-controlled solvent delivery system. The reaction progress and completion can be monitored in real time using a standard UV detector. All steps of oligosaccharide assembly including loading, glycosylation, deprotection, and cleavage can be performed using this setup.     

NanoPARCEL: a method for controlling cellular behavior with external light

We developed a simple preparation procedure for the protein encapsulated nanoparticle and used the nanoparticle for spatiotemporal activity control of various proteins. We succeeded in the local protein activation within cells by light using the nanoparticle.     

Controlled synthesis of hydroxyapatite-supported palladium complexes as highly efficient heterogeneous catalysts

Achieving precise control of active species on solid surfaces is one of the most important goals in the development of highly functionalized heterogeneous catalysts. The treatment of hydroxyapatites with PdCl(2)(PhCN)(2) gives two new types of hydroxyapatite-bound Pd complexes. Using the stoichiometric hydroxyapatite, Ca(10)(PO(4))(6)(OH)(2), we found that monomeric PdCl(2) species can be grafted on its surface, which are easily transformed into Pd(0) particles with narrow size distribution in the presence of alcohols. Such metallic Pd species can effectively promote alcohol oxidation using molecular oxygen and are shown to give a remarkably high TON of up to 236 000. Another monomeric Pd(II) phosphate complex can be generated at a Ca-deficient site of the nonstoichiometric hydroxyapatite, Ca(9)(HPO(4))(PO(4))(5)(OH), affording a catalyst with Pd(II) structure and high activity for the Heck and Suzuki reactions. To the best of our knowledge, the PdHAP are one of the most active heterogeneous catalysts for both alcohol oxidation under an atmospheric O(2)() pressure and the Heck reaction reported to date. These Pd catalysts are recyclable in the above organic reactions. Our approach to catalyst preparation based on the control of Ca/P ratios of hydroxyapatites represents a particularly attractive method for the nanoscale design of catalysts.     

Highly efficient esterification of carboxylic acids with alcohols by montmorillonite-enwrapped titanium as a heterogeneous acid catalyst

Montmorillonite-enwrapped titanium catalyst was found to efficiently promote the esterification of carboxylic acids with alcohols. In comparison to other catalysts reported to date, this heterogeneous catalyst offers a remarkably simple workup procedure, and is reusable without any appreciable loss in its activity and selectivity.