Crystal structures of monkey and mouse nicotinamide N-methyltransferase (NNMT) bound with end product, 1-methyl nicotinamide

Birudukota, Swarnakumari; Thakur, Manish K.; Parveen, R.; Kandan, Saravanan; Joghee, Suresh; Shaik, Shama; Anand, Niranjan Naranapura; Burri, Raghunadha Reddy; Kristam, Rajendra; Hallur, Mahanandeesha Siddappa; Rajagopal, Sridharan; Schreuder, Herman; Langer, Thomas; Rudolph, Christine; Ruf, Sven; Dhakshinamoorthy, Saravanakumar; Gosu, Ramachandraiah; Kannt, Aimo

doi:10.1016/j.bbrc.2017.07.087

Cited by 10 publications

(13 citation statements)

References 14 publications

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“…During the preparation of this manuscript, crystal structures of monkey and mouse NNMT in complex with 1 and SAH were published. 43 Our hNNMT−6 structure overlays very well with these two new structures with almost identical ligandprotein interactions (Figure 6D and Supporting Information Figure S5). Interestingly, while the 3-amido phenyl group of compound 6 is located deeper in the substrate binding site compared to nicotinamide, this group sits at almost the same location as 1 in the monkey NNMT−1 complex (Figure 6C,D).…”

Section: ■ Results and Discussionsupporting

confidence: 63%

Discovery of Bisubstrate Inhibitors of Nicotinamide N-Methyltransferase (NNMT)

Babault

Allali-Hassani

et al. 2018

J. Med. Chem.

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Nicotinamide N-methyltransferase (NNMT) catalyzes the N-methylation of pyridine-containing compounds using the cofactor S-5'-adenosyl-l-methionine (SAM) as the methyl group donor. Through the regulation of the levels of its substrates, cofactor, and products, NNMT plays an important role in physiology and pathophysiology. Overexpression of NNMT has been implicated in various human diseases. Potent and selective small-molecule NNMT inhibitors are valuable chemical tools for testing biological and therapeutic hypotheses. However, very few NNMT inhibitors have been reported. Here, we describe the discovery of a bisubstrate NNMT inhibitor MS2734 (6) and characterization of this inhibitor in biochemical, biophysical, kinetic, and structural studies. Importantly, we obtained the first crystal structure of human NNMT in complex with a small-molecule inhibitor. The structure of the NNMT-6 complex has unambiguously demonstrated that 6 occupied both substrate and cofactor binding sites. The findings paved the way for developing more potent and selective NNMT inhibitors in the future.

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Section: ■ Results and Discussionsupporting

confidence: 63%

Discovery of Bisubstrate Inhibitors of Nicotinamide N-Methyltransferase (NNMT)

Babault

Allali-Hassani

et al. 2018

J. Med. Chem.

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“…The full length NNMT proteins (1–264 amino acids) of human & mouse (UniProt accession # P40261 & O55239) with an amino terminal 6 × His tag and TEV cleavage site (ENLYFQG) were cloned into pET23a between NdeI and XhoI restriction sites and expressed in E. coli Rosetta2(DE3)pLysS cells. The recombinant proteins were purified as described previously 24 .…”

Section: Methodsmentioning

confidence: 99%

“…Three point mutations were introduced in human NNMT (K100A/E101A/E103A) using the QuikChange kit (Stratagene). The proteins were produced and purified from E. coli as previously described 24 . Final purified human NNMT sample was concentrated to 12 mg/mL in buffer (20 mM HEPES pH 7.5, 50 mM NaCl, 5% (v/v) glycerol, 1 mM TCEP) and mouse NNMT sample was concentrated to 12 mg/mL in buffer (20 mM HEPES pH 7.5, 125 mM NaCl, 5% (v/v) glycerol, 1 mM TCEP)…”

Section: Methodsmentioning

confidence: 99%

A small molecule inhibitor of Nicotinamide N-methyltransferase for the treatment of metabolic disorders

Kannt

Rajagopal

Kadnur

et al. 2018

Sci Rep

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101

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Nicotinamide N-methyltransferase (NNMT) is a cytosolic enzyme that catalyzes the transfer of a methyl group from the co-factor S-adenosyl-L-methionine (SAM) onto the substrate, nicotinamide (NA) to form 1-methyl-nicotinamide (MNA). Higher NNMT expression and MNA concentrations have been associated with obesity and type-2 diabetes. Here we report a small molecule analog of NA, JBSNF-000088, that inhibits NNMT activity, reduces MNA levels and drives insulin sensitization, glucose modulation and body weight reduction in animal models of metabolic disease. In mice with high fat diet (HFD)-induced obesity, JBSNF-000088 treatment caused a reduction in body weight, improved insulin sensitivity and normalized glucose tolerance to the level of lean control mice. These effects were not seen in NNMT knockout mice on HFD, confirming specificity of JBSNF-000088. The compound also improved glucose handling in ob/ob and db/db mice albeit to a lesser extent and in the absence of weight loss. Co-crystal structure analysis revealed the presence of the N-methylated product of JBSNF-000088 bound to the NNMT protein. The N-methylated product was also detected in the plasma of mice treated with JBSNF-000088. Hence, JBSNF-000088 may act as a slow-turnover substrate analog, driving the observed metabolic benefits.

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“…Byproducts of NNMT, SAH and 1-MNA inhibit NNMT activity via a feedback mechanism ( 107 ). A study of comprehensive structure–activity relationships (SARs) revealed that quinolinium analogues inhibit NNMT at low concentrations and selectively bind to the NNMT substrate-binding site residues ( 99 ).…”

Section: Development and Application Of Nnmt In Cancer Therapymentioning

confidence: 99%

Nicotinamide N-Methyltransferase: A Promising Biomarker and Target for Human Cancer Therapy

Chen

et al. 2022

Front. Oncol.

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Cancer cells typically exhibit a tightly regulated program of metabolic plasticity and epigenetic remodeling to meet the demand of uncontrolled cell proliferation. The metabolic–epigenetic axis has recently become an increasingly hot topic in carcinogenesis and offers new avenues for innovative and personalized cancer treatment strategies. NicotinamideN-methyltransferase (NNMT) is a metabolic enzyme involved in controlling methylation potential, impacting DNA and histone epigenetic modification.NNMToverexpression has been described in various solid cancer tissues and even body fluids, including serum, urine, and saliva. Furthermore, accumulating evidence has shown thatNNMTknockdown significantly decreases tumorigenesis and chemoresistance capacity. Most importantly, the natural NNMT inhibitor yuanhuadine can reverse epidermal growth factor receptor tyrosine kinase inhibitor resistance in lung cancer cells. In this review, we evaluate the possibility of NNMT as a diagnostic biomarker and molecular target for effective anticancer treatment. We also reveal the exact mechanisms of how NNMT affects epigenetics and the development of more potent and selective inhibitors.

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Crystal structures of monkey and mouse nicotinamide N-methyltransferase (NNMT) bound with end product, 1-methyl nicotinamide

Cited by 10 publications

References 14 publications

Discovery of Bisubstrate Inhibitors of Nicotinamide N-Methyltransferase (NNMT)

Discovery of Bisubstrate Inhibitors of Nicotinamide N-Methyltransferase (NNMT)

A small molecule inhibitor of Nicotinamide N-methyltransferase for the treatment of metabolic disorders

Nicotinamide N-Methyltransferase: A Promising Biomarker and Target for Human Cancer Therapy

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