Abstract
Nicotinamide adenine dinucleotide (NAD) kinases are essential and ubiquitous enzymes involved in the tight regulation of NAD/nicotinamide adenine dinucleotide phosphate (NADP) levels in many metabolic pathways. Consequently, they represent promising therapeutic targets in cancer and antibacterial treatments. We previously reported diadenosine derivatives as NAD kinase inhibitors with bactericidal activities on Staphylococcus aureus. Among them, one compound (namely NKI1) was found effective in vivo in a mouse infection model. With the aim to gain detailed knowledge about the selectivity and mechanism of action of this lead compound, we planned to develop a chemical probe that could be used in affinity-based chemoproteomic approaches. Here, we describe the first functionalized chemical probe targeting a bacterial NAD kinase. Aminoalkyl functional groups were introduced on NKI1 for further covalent coupling to an activated SepharoseTM matrix. Inhibitory properties of functionalized NKI1 derivatives together with X-ray characterization of their complexes with the NAD kinase led to identify candidate compounds that are amenable to covalent coupling to a matrix.
Highlights
Nicotinamide adenine dinucleotide (NAD) kinases (NADK) are ubiquitous enzymes, which catalyze the phosphorylation of NAD to nicotinamide adenine dinucleotide phosphate (NADP), which is subsequently reduced to NADPH [1,2,3]
Following a fragment-based approach, we identified a series of diadenosine derivatives with low micromolar inhibitory potencies against recombinant LmNADK1 and S. aureus NADK [16,17]
We subsequently discovered the first NAD kinase inhibitor active in mice infected with S. aureus, including antibiotic-resistant strains [18]
Summary
Nicotinamide adenine dinucleotide (NAD) kinases (NADK) are ubiquitous enzymes, which catalyze the phosphorylation of NAD to nicotinamide adenine dinucleotide phosphate (NADP), which is subsequently reduced to NADPH [1,2,3]. Since it is the only known enzyme producing NADP de novo, NAD kinase plays a crucial role in controlling the intracellular balance of NAD(H) and NADP(H) in many cellular metabolic pathways [4,5]. While the NADK enzymatic activity has been known for decades, their genes were identified more recently, leading to the discovery of orthologs in most living organisms. It was shown that human NAD kinase displays kinetic and structural features that differ considerably from that of prokaryotes [13,14].
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