Structure of arylamineN-acetyltransferase fromMycobacterium tuberculosisdetermined by cross-seeding with the homologous protein fromM. marinum: triumph over adversity

Abuhammad, Areej; Lowe, E.D.; McDonough, Michael A.; Stewart, Patrick D. Shaw; Kolek, Stefan; Sim, Edith; Garman, Elspeth F.

doi:10.1107/s0907444913015126

Cited by 27 publications

(39 citation statements)

References 86 publications

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“…While we were improving the yield of the TBNAT enzyme, we used NAT from M. smegmatis (Sandy et al ., ) and then M. marinum (Fullam et al ., ), which is much closer in sequence to the NAT from M. tuberculosis . M. marinum NAT is very soluble while the M. tuberculosis NAT is not (Fullam et al ., ; Abuhammad et al ., in press) and they show very different physical properties (Lack et al ., ), including melting at different temperatures. The melting property resides in the first two domains of the protein and may reflect the ability of TBNAT to remain active in inflammatory conditions associated with intracellular infection.…”

Section: Knockout Micementioning

confidence: 99%

“…We have at last, after 15 years, obtained a crystal structure of the NAT from M. tuberculosis by seeding a preparation of M. tuberculosis NAT with a crystal of NAT from M. marinum (Abuhammad et al, 2013). The substrate specificities of the M. marinum and M. tuberculosis NAT enzymes are not identical and this illustrates the importance in drug discovery of using an appropriate model or at least understanding the differences when a surrogate has to be used.…”

Section: Nat In Bacteriamentioning

confidence: 99%

See 1 more Smart Citation

Arylamine N‐acetyltransferases: from drug metabolism and pharmacogenetics to drug discovery

Abuhammad

Ryan

2014

British J Pharmacology

131

105

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Arylamine N-acetyltransferases (NATs) are polymorphic drug-metabolizing enzymes, acetylating arylamine carcinogens and drugs including hydralazine and sulphonamides. The slow NAT phenotype increases susceptibility to hydralazine and isoniazid toxicity and to occupational bladder cancer. The two polymorphic human NAT loci show linkage disequilibrium. All mammalian Nat genes have an intronless open reading frame and non-coding exons. The human gene products NAT1 and NAT2 have distinct substrate specificities: NAT2 acetylates hydralazine and human NAT1 acetylates p-aminosalicylate (p-AS) and the folate catabolite para-aminobenzoylglutamate (p-abaglu). Human NAT2 is mainly in liver and gut. Human NAT1 and its murine homologue are in many adult tissues and in early embryos. Human NAT1 is strongly expressed in oestrogen receptor-positive breast cancer and may contribute to folate and acetyl CoA homeostasis. NAT enzymes act through a catalytic triad of Cys, His and Asp with the architecture of the active site-modulating specificity. Polymorphisms may cause unfolded protein. The C-terminus helps bind acetyl CoA and differs among NATs including prokaryotic homologues. NAT in Salmonella typhimurium supports carcinogen activation and NAT in mycobacteria metabolizes isoniazid with polymorphism a minor factor in isoniazid resistance. Importantly, nat is in a gene cluster essential for Mycobacterium tuberculosis survival inside macrophages. NAT inhibitors are a starting point for novel anti-tuberculosis drugs. Human NAT1-specific inhibitors may act in biomarker detection in breast cancer and in cancer therapy. NAT inhibitors for co-administration with 5-aminosalicylate (5-AS) in inflammatory bowel disease has prompted ongoing investigations of azoreductases in gut bacteria which release 5-AS from prodrugs including balsalazide.

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Section: Knockout Micementioning

confidence: 99%

Section: Nat In Bacteriamentioning

confidence: 99%

Arylamine N‐acetyltransferases: from drug metabolism and pharmacogenetics to drug discovery

Abuhammad

Ryan

2014

British J Pharmacology

131

105

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“…The obtained protein sequences of MTB and human were submitted to protein alignment program (BLASTp)19 and searched against protein database (PDB; http://www.rcsb.org/pdb/home/home.do). The WT-NAT and NAT2 protein structures of MTB and human (4BGF and 2PFR) were considered as the templates, respectively2021.…”

Section: Methodsmentioning

confidence: 99%

“…In other words, the MT of NAT of MTB (G207R) was generated using crystal structure (4BGF) of NAT protein of MTB obtained from PDB20. In case of human the MT of NAT2 (K268R) was generated using crystal structure 2PFR, WT-NAT2 protein of human21.…”

Section: Methodsmentioning

confidence: 99%

The Role ofN-Acetyl Transferases on Isoniazid Resistance fromMycobacterium tuberculosisand Human: AnIn SilicoApproach

2017

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BackgroundN-acetyl transferase (NAT) inactivates the pro-drug isoniazid (INH) to N-acetyl INH through a process of acetylation, and confers low-level resistance to INH in Mycobacterium tuberculosis (MTB). Similar to NAT of MTB, NAT2 in humans performs the same function of acetylation. Rapid acetylators, may not respond to INH treatment efficiently, and could be a potential risk factor, for the development of INH resistance in humans.MethodsTo understand the contribution of NAT of MTB and NAT2 of humans in developing INH resistance using in silico approaches, in this study, the wild type (WT) and mutant (MT)-NATs of MTB, and humans, were modeled and docked, with substrates and product (acetyl CoA, INH, and acetyl INH). The MT models were built, using templates 4BGF of MTB, and 2PFR of humans.ResultsOn the basis of docking results of MTB-NAT, it can be suggested that in comparison to the WT, binding affinity of MT-G207R, was found to be lower with acetyl CoA, and higher with acetyl-INH and INH. In case of MT-NAT2 from humans, the pattern of score with respect to acetyl CoA and acetyl-INH, was similar to MT-NAT of MTB, but revealed a decrease in INH score.ConclusionIn MTB, MT-NAT revealed high affinity towards acetyl-INH, which can be interpreted as increased formation of acetyl-INH, and therefore, may lead to INH resistance through inactivation of INH. Similarly, in MT-NAT2 (rapid acetylators), acetylation occurs rapidly, serving as a possible risk factor for developing INH resistance in humans.

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“…NAT2 and TBNAT structural models were obtained from crystallographic structures deposited in the Protein Data Bank [39,40] (PDB) (www.rcsb.org): NAT2, accession code 2PFR (resolution 1.9 Å) [50] and TBNAT, accession code 4BGF (resolution 2.1 Å) [51]. Since the two human NAT1 structures deposited in PDB have missing atoms and modified amino acid residues, the structural model of NAT1 was obtained by homology modelling.…”

Section: Target Preparationmentioning

confidence: 99%

In Silico Screening and Analysis of Potential Inhibitors of Arylamine N-Acetyltransferases (NATs) from the Traditional Chinese Medicine: A Study Using Free Available Tools

Azevedo¹,

Richardt²,

Oliveira³

et al. 2017

Preprint

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Arylamine N-acetyltransferases (NATs) are cytosolic enzymes, highly polymorphic, present in both eukaryotes and prokaryotes. These enzymes play an important role in the detoxification and activation of xenobiotics as well as in the synthesis of endogenous compounds. Specific NATs have been pointed out in the literature as possible therapeutic targets. In particular, the human NAT1, for the treatment of certain cancers, and the NAT from M. tuberculosis (TBNAT), for the treatment of tuberculosis. This paper describes an in silico approach to prospect and select potentially inhibitors of NAT1 and TBNAT from the Traditional Chinese Medicine (TCM) using free available tools. A library with ligands from TCM was previously screened in order to select only compounds with optimal pharmacological properties. The affinity of the selected ligands with respect to NAT enzymes was then evaluated by virtual screening (VS). Subsequently, the complexes with the best ligands were submitted to molecular dynamics (MD) simulations aiming to obtain better quality information on affinity and selectivity. The results for one specific ligand, ZINC14690579, indicated its potential for affinity and selectivity. ZINC14690579 structure may represent the discovery of a new scaffold for future development of NAT inhibitors.

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Structure of arylamineN-acetyltransferase fromMycobacterium tuberculosisdetermined by cross-seeding with the homologous protein fromM. marinum: triumph over adversity

Cited by 27 publications

References 86 publications

Arylamine N‐acetyltransferases: from drug metabolism and pharmacogenetics to drug discovery

Arylamine N‐acetyltransferases: from drug metabolism and pharmacogenetics to drug discovery

The Role ofN-Acetyl Transferases on Isoniazid Resistance fromMycobacterium tuberculosisand Human: AnIn SilicoApproach

<em>In Silico</em> Screening and Analysis of Potential Inhibitors of Arylamine N-Acetyltransferases (NATs) from the Traditional Chinese Medicine: A Study Using Free Available Tools

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