Preprint Article Version 1 This version is not peer-reviewed

Membrane-Permeable Octanoyloxybenzyl-Masked cNMPs as Novel Tools for Non-Invasive cell Assays

Version 1 : Received: 12 October 2018 / Approved: 12 October 2018 / Online: 12 October 2018 (11:32:45 CEST)

A peer-reviewed article of this Preprint also exists.

Ruthenbeck, A.; Marangoni, E.; Diercks, B.-P.; Krüger, A.; Froese, A.; Bork, N.I.; Nikolaev, V.O.; Guse, A.H.; Meier, C. Membrane-Permeable Octanoyloxybenzyl-Masked cNMPs As Novel Tools for Non-Invasive Cell Assays. Molecules 2018, 23, 2960. Ruthenbeck, A.; Marangoni, E.; Diercks, B.-P.; Krüger, A.; Froese, A.; Bork, N.I.; Nikolaev, V.O.; Guse, A.H.; Meier, C. Membrane-Permeable Octanoyloxybenzyl-Masked cNMPs As Novel Tools for Non-Invasive Cell Assays. Molecules 2018, 23, 2960.

Journal reference: Molecules 2018, 23, 2960
DOI: 10.3390/molecules23112960

Abstract

Adenine nucleotide (AN) 2nd messengers such as 3’,5’-cyclic adenosine monophosphate (cAMP) are central elements of intracellular signaling, but many details of underlying processes remain still elusive. Like all nucleotides, cyclic nucleotide monophosphates (cNMPs) are net-negatively charged at physiologic pH which limits their applicability in cell-based settings. Thus, many cellular assays rely on sophisticated techniques like microinjection or electroporation. This setup is not feasible for medium- to high-throughput formats, and the mechanic stress that cells are exposed to raises the probability of interfering artefacts or false-positives. Here, we present a short and flexible chemical route yielding membrane-permeable, bio-reversibly masked cNMPs for which we employed the octanoyloxybenzyl (OB) group. We further show hydrolysis studies on chemical stability and enzymatic activation, and present results of real-time assays, where we used cAMP and Ca2+ live cell imaging to demonstrate high permeability and prompt intracellular conversion of some selected masked cNMPs. Consequently, our novel OB-masked cNMPs constitute valuable precursor-tools for non-invasive studies on intracellular signaling.

Subject Areas

cyclic nucleotide monophosphate; bio-reversible protection; acyloxybenzyl phosphate ester

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