5NSD

Co-crystal structure of NAMPT dimer with KPT-9274

Summary for 5NSD

• Entry DOI: 10.2210/pdb5nsd/pdb
• Descriptor: Nicotinamide phosphoribosyltransferase, PHOSPHATE ION, GLYCEROL, ... (5 entities in total)
• Functional Keywords: drug target, nampt, crispr/cas, transferase
• Biological source: Homo sapiens (Human)
• Cellular location: Nucleus : P43490
• Total number of polymer chains: 2
• Total formula weight: 115480.08

Authors

Neggers, J.E.,Kwanten, B.,Dierckx, T.,Noguchi, H.,Voet, A.,Vercruysse, T.,Baloglu, E.,Senapedis, W.,Jacquemyn, M.,Daelemans, D. (deposition date: 2017-04-26, release date: 2018-02-14, Last modification date: 2024-05-08)

Primary citation

Neggers, J.E.,Kwanten, B.,Dierckx, T.,Noguchi, H.,Voet, A.,Bral, L.,Minner, K.,Massant, B.,Kint, N.,Delforge, M.,Vercruysse, T.,Baloglu, E.,Senapedis, W.,Jacquemyn, M.,Daelemans, D.
Target identification of small molecules using large-scale CRISPR-Cas mutagenesis scanning of essential genes.
Nat Commun, 9:502-502, 2018

PubMed Abstract: Unraveling the mechanism of action and molecular target of small molecules remains a major challenge in drug discovery. While many cancer drugs target genetic vulnerabilities, loss-of-function screens fail to identify essential genes in drug mechanism of action. Here, we report CRISPRres, a CRISPR-Cas-based genetic screening approach to rapidly derive and identify drug resistance mutations in essential genes. It exploits the local genetic variation created by CRISPR-Cas-induced non-homologous end-joining (NHEJ) repair to generate a wide variety of functional in-frame mutations. Using large sgRNA tiling libraries and known drug-target pairs, we validate it as a target identification approach. We apply CRISPRres to the anticancer agent KPT-9274 and identify nicotinamide phosphoribosyltransferase (NAMPT) as its main target. These results present a powerful and simple genetic approach to create many protein variants that, in combination with positive selection, can be applied to reveal the cellular target of small-molecule inhibitors.

PubMed: 29402884
DOI: 10.1038/s41467-017-02349-8

Experimental method

X-RAY DIFFRACTION (2.046 Å)