A Modular Synthesis of Modified Phosphoanhydrides

Abstract

Phosphoanhydrides (P-anhydrides) are ubiquitously occurring modifications in nature. Nucleotides and their conjugates, for example, are among the most important building blocks and signaling molecules in cell biology. To study and manipulate their biological functions, a diverse range of analogues have been developed. Phosphate-modified analogues have been successfully applied to study proteins that depend on these abundant cellular building blocks, but very often both the preparation and purification of these molecules are challenging. This study discloses a general access to P-anhydrides, including different nucleotide probes, that greatly facilitates their preparation and isolation. The convenient and scalable synthesis of, for example, (18) O labeled nucleoside triphosphates holds promise for future applications in phosphoproteomics.

Identifying Kinase Substrates via a Heavy ATP Kinase Assay and Quantitative Mass Spectrometry

“…Beads were washed 2× with IP-buffer and 1× with KA buffer (40 mM Tris-HCl pH 7.5, 10 mM MgCl 2 , 1 mM DTT), pooled and resuspended in KA buffer. The beads were equally split into eppendorf tubes, 5 mg FSBA-treated cellular protein fraction added and the kinase reaction initiated by adding ‘light’ (SIGMA-Aldrich) or ‘heavy’ 18 O 2 -ATP (Organisch-Chemisches Institut, UZH, Switzerland)29 at a final concentration of 1 mM. After incubation for 30, 90 and 150 min at RT (~23 °C) on a rotary wheel, the reaction was quenched by the addition of solid urea to a final concentration of 8 M. Samples were vortexed and stored on ice.…”

Chemical ADP-ribosylation: mono-ADPr-peptides and oligo-ADP-ribose

“…Pyrophosphates are important functional groups in a wide array of naturally occurring compounds and a lot of procedures for the synthesis of pyrophosphates have been reported. 19,[40][41][42][43][44][45][46][47][48] However, the occurrence of multiple pyrophosphates in one molecule, such as in oligo-ADPr, is unprecedented and presents a special challenge. Only two syntheses of short fragments of oligo-ADPr have been published to date.…”

Paracelsus‐Preis: M. Grätzel / Werner‐Preis: M. Kovalenko / Ružička‐Preis: H. J. Jessen / Österreichisches Ehrenkreuz für Wissenschaft und Kunst 1. Klasse: B. Kräutler

“…Im Mittelpunkt seiner Forschung steht die Synthese hochphosphorylierter sekundärer Botenstoffe, ihre Modifikation und ihr Einsatz in chemisch‐biologischen Studien. Von ihm erschien in Chemistry—A European Journal eine Arbeit über die modulare Synthese modifizierter Phosphoanhydride und in der Angewandten Chemie eine über Diphospho‐ myo ‐inositolphosphate …”