Chemical Synthesis and Enzymatic Testing of CMP‐Sialic Acid Derivatives

The cycloSal approach has been used in the past for the synthesis of a range of phosphorylated bioconjugates. In those reports, cycloSal nucleotides were allowed to react with different phosphate nucleophiles. With glycopyranosyl phosphates as nucleophiles, diphosphate‐linked sugar nucleotides were formed. Here, cycloSal‐nucleotides were used to prepare monophosphate‐linked sugar nucleotides successfully in high anomeric purity and high chemical yield. The method was successfully used for the synthesis of three nucleotide glycopyranoses as model compounds. The method was then applied to the syntheses of CMP‐N‐acetyl‐neuraminic acids (CMP‐Neu5NAc) and of four derivatives with different modifications at their amino functions (N‐propanoyl, N‐butanoyl, N‐pentanoyl and N‐cyclopropylcarbonyl). The compounds were used for initial enzymatic studies with a bacterial polysialyltransferase (polyST). Surprisingly, the enzyme showed marked differences in terms of utilisation of the four derivatives. The N‐propanoyl, N‐butanoyl, and N‐pentanoyl derivatives were efficiently used in a first transfer with a fluorescently labelled trisialo‐acceptor. However, elongation of the resulting tetrasialo‐acceptors worsened progressively with the size of the N‐acyl chain. The N‐pentanoyl derivative allowed a single transfer, leading to a capped tetramer. The N‐cyclopropylcarbonyl derivative was not transferred.     

Quantitative Imaging of the Magnetic Configuration of Modulated Nanostructures by Electron Holography

By means of off‐axis electron holography the local distribution of the magnetic induction within and around a poly‐crystalline Permalloy (Ni₈₁Fe₁₉) thin film is studied. In addition the stray field above the sample is measured by magnetic force microscopy on a larger area. The film is deposited on a periodically nanostructured (rippled) Si substrate, which was formed by Xe⁺ ion beam erosion. This introduces the periodical ripple shape to the Permalloy film. The created ripple morphology is expected to modify the magnetization distribution within the Permalloy and to induce dipolar stray fields. These stray fields play an important role in spinwave dynamics of periodic nanostructures like magnonic crystals. Micromagnetic simulations estimate those stray fields in the order of only 10 mT. Consequently, their experimental determination at nanometer spatial resolution is highly demanding and requires advanced acquisition and reconstruction techniques such as electron holography. The reconstructed magnetic phase images show the magnetized thin film, in which the magnetization direction follows mainly the given morphology. Furthermore, a closer look to the Permalloy/carbon interface reveals stray fields at the detection limit of the method in the order of 10 mT, which is in qualitative agreement with the micromagnetic simulations.