scholarly journals Mycobacterial Trehalose Synthase as a potential drug target for tuberculosis

2014 ◽  
Vol 70 (a1) ◽  
pp. C441-C441
Author(s):  
Sami Caner ◽  
Nham Nguyen ◽  
Adeleke Aguda ◽  
Ran Zhang ◽  
Yuan Pan ◽  
...  
Keyword(s):  
Active Site ◽  
Mycobacterium Smegmatis ◽  
Drug Target ◽  
The Other ◽  
Intramolecular Rearrangement ◽  
Biosynthetic Pathways ◽  
Potential Drug Target ◽  
Trehalose Synthase ◽  
Active Site Cleft ◽  
Glycogen Biosynthesis

Trehalose synthase (TreS) catalyzes the reversible conversion of maltose to trehalose in mycobacteria as one of three biosynthetic pathways to this non-reducing disaccharide. Given the importance of trehalose to survival of mycobacteria there has been considerable interest in understanding the enzymes involved in its production; indeed the structures of the key enzymes in the other two pathways have already been determined. Herein we present the first structure of TreS from Mycobacterium smegmatis, thereby providing insights into the catalytic machinery involved in this intriguing intramolecular reaction. This structure, which is of interest both mechanistically and as a potential pharmaceutical target, reveals a narrow and enclosed active site cleft within which the intramolecular rearrangement can occur with minimal hydrolysis. We also present the structure of a complex of TreS with acarbose, revealing a hitherto unsuspected oligosaccharide binding site within the C-terminal domain. This may well provide an anchor point for the association of TreS with glycogen, thereby enhancing its role in glycogen biosynthesis and degradation.

Active site acidic residues and structural analysis of modelled human aromatase: A potential drug target for breast cancer

2006 ◽  
Vol 19 (12) ◽  
pp. 857-870 ◽  
Author(s):  
J. Narashima Murthy ◽  
M. Nagaraju ◽  
G. Madhavi Sastry ◽  
A. Raghuram Rao ◽  
G.␣Narahari Sastry
Keyword(s):  
Breast Cancer ◽  
Structural Analysis ◽  
Active Site ◽  
Drug Target ◽  
Potential Drug Target ◽  
Potential Drug ◽  
Acidic Residues ◽  
Human Aromatase

scholarly journals Chemical validation of Mycobacterium tuberculosis phosphopantetheine adenylyltransferase using fragment linking and CRISPR interference

2020 ◽  
Author(s):  
Jamal El Bakali ◽  
Michal Blaszczyk ◽  
Joanna C. Evans ◽  
Jennifer A. Boland ◽  
William J. McCarthy ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Crystal Structures ◽  
Active Site ◽  
Drug Target ◽  
Potential Drug Target ◽  
Biosynthesis Pathway ◽  
Potential Drug ◽  
Antimicrobial Drugs ◽  
X Ray ◽  
Crispr Interference

AbstractThe coenzyme A (CoA) biosynthesis pathway has attracted attention as a potential target for much-needed novel antimicrobial drugs, including for the treatment of tuberculosis (TB), the lethal disease caused by Mycobacterium tuberculosis (Mtb). Seeking to identify the first inhibitors of Mtb phosphopantetheine adenylyltransferase (MtbPPAT), the enzyme that catalyses the penultimate step in CoA biosynthesis, we performed a fragment screen. In doing so, we discovered three series of fragments that occupy distinct regions of the MtbPPAT active site, presenting a unique opportunity for fragment linking. Here we show how, guided by X-ray crystal structures, we could link weakly-binding fragments to produce an active site binder with a KD < 20 μM and on-target anti-Mtb activity, as demonstrated using CRISPR interference. This study represents a big step toward validating MtbPPAT as a potential drug target and designing a MtbPPAT-targeting anti-TB drug.Abstract Figure

scholarly journals A structure of substrate-bound Synaptojanin1 provides new insights in its mechanism and the effect of disease mutations

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Jone Paesmans ◽  
Ella Martin ◽  
Babette Deckers ◽  
Marjolijn Berghmans ◽  
Ritika Sethi ◽  
...  
Keyword(s):  
Active Site ◽  
Drug Target ◽  
Molecular Mechanisms ◽  
Potential Drug Target ◽  
Missense Mutations ◽  
Loss Of Function ◽  
Phosphatase Domain ◽  
Phosphoinositide Phosphatase ◽  
Disease Mutations ◽  
Phosphate Groups

Synaptojanin1 (Synj1) is a phosphoinositide phosphatase, important in clathrin uncoating during endocytosis of presynaptic vesicles. It was identified as a potential drug target for Alzheimer’s disease, Down syndrome, and TBC1D24-associated epilepsy, while also loss-of-function mutations in Synj1 are associated with epilepsy and Parkinson’s disease. Despite its involvement in a range of disorders, structural, and detailed mechanistic information regarding the enzyme is lacking. Here, we report the crystal structure of the 5-phosphatase domain of Synj1. Moreover, we also present a structure of this domain bound to the substrate diC8-PI(3,4,5)P3, providing the first image of a 5-phosphatase with a trapped substrate in its active site. Together with an analysis of the contribution of the different inositide phosphate groups to catalysis, these structures provide new insights in the Synj1 mechanism. Finally, we analysed the effect of three clinical missense mutations (Y793C, R800C, Y849C) on catalysis, unveiling the molecular mechanisms underlying Synj1-associated disease.

scholarly journals Structure of the Bacillus anthracis dTDP-L-rhamnose-biosynthetic enzyme dTDP-4-dehydrorhamnose 3,5-epimerase (RfbC)

2017 ◽  
Vol 73 (12) ◽  
pp. 664-671 ◽  
Author(s):  
Aleksander Shornikov ◽  
Ha Tran ◽  
Jennifer Macias ◽  
Andrei S. Halavaty ◽  
George Minasov ◽  
...  
Keyword(s):  
Bacillus Anthracis ◽  
Immune Evasion ◽  
Active Site ◽  
Drug Target ◽  
Biosynthetic Pathway ◽  
The Other ◽  
Biosynthetic Enzyme ◽  
Active Site Residues ◽  
Structural Comparison ◽  
Cell Wall Component

The exosporium layer of Bacillus anthracis spores is rich in L-rhamnose, a common bacterial cell-wall component, which often contributes to the virulence of pathogens by increasing their adherence and immune evasion. The biosynthetic pathway used to form the activated L-rhamnose donor dTDP-L-rhamnose consists of four enzymes (RfbA, RfbB, RfbC and RfbD) and is an attractive drug target because there are no homologs in mammals. It was found that co-purifying and screening RfbC (dTDP-6-deoxy-D-xylo-4-hexulose 3,5-epimerase) from B. anthracis in the presence of the other three B. anthracis enzymes of the biosynthetic pathway yielded crystals that were suitable for data collection. RfbC crystallized as a dimer and its structure was determined at 1.63 Å resolution. Two different ligands were bound in the protein structure: pyrophosphate in the active site of one monomer and dTDP in the other monomer. A structural comparison with RfbC homologs showed that the key active-site residues are conserved across kingdoms.

scholarly journals An Enlarged, Adaptable Active Site in CYP164 Family P450 Enzymes, the Sole P450 in Mycobacterium leprae

2011 ◽  
Vol 56 (1) ◽  
pp. 391-402 ◽  
Author(s):  
Christopher R. J. Agnew ◽  
Andrew G. S. Warrilow ◽  
Nicholas M. Burton ◽  
David C. Lamb ◽  
Steven L. Kelly ◽  
...  
Keyword(s):  
Active Site ◽  
Mycobacterium Smegmatis ◽  
Metal Ion ◽  
Drug Target ◽  
Mycobacterium Leprae ◽  
Homology Model ◽  
Binding Studies ◽  
Content Type ◽  
Computational Docking ◽  
Primary Location

ABSTRACTCYP164 family P450 enzymes are found in only a subset of mycobacteria and include CYP164A1, which is the sole P450 found inMycobacterium leprae, the causative agent of leprosy. This has previously led to interest in this enzyme as a potential drug target. Here we describe the first crystal structure of a CYP164 enzyme, CYP164A2 fromMycobacterium smegmatis. CYP164A2 has a distinctive, enlarged hydrophobic active site that extends above the porphyrin ring toward the access channels. Unusually, we find that CYP164A2 can simultaneously bind two econazole molecules in different regions of the enlarged active site and is accompanied by the rearrangement and ordering of the BC loop. The primary location is through a classic interaction of the azole group with the porphyrin iron. The second econazole molecule is bound to a unique site and is linked to a tetracoordinated metal ion complexed to one of the heme carboxylates and to the side chains of His 105 and His 364. All of these features are preserved in the closely homologousM. lepraeCYP164A1. The computational docking of azole compounds to a homology model of CYP164A1 suggests that these compounds will form effective inhibitors and is supported by the correlation of parallel docking with experimental binding studies of CYP164A2. The binding of econazole to CYP164A2 occurs primarily through the high-spin “open” conformation of the enzyme (Kd[dissociation constant] of 0.1 μM), with binding to the low-spin “closed” form being significantly hindered (Kdof 338 μM). These studies support previous suggestions that azole derivatives may provide an effective strategy to improve the treatment of leprosy.

scholarly journals The structure of the Mycobacterium smegmatis trehalose synthase reveals an unusual active site configuration and acarbose-binding mode

Glycobiology ◽  
2013 ◽  
Vol 23 (9) ◽  
pp. 1075-1083 ◽  
Author(s):  
S. Caner ◽  
N. Nguyen ◽  
A. Aguda ◽  
R. Zhang ◽  
Y. T. Pan ◽  
...  
Keyword(s):  
Active Site ◽  
Mycobacterium Smegmatis ◽  
Binding Mode ◽  
Trehalose Synthase

Inhibition of 7,8-diaminopelargonic acid aminotransferase by amiclenomycin and analogues

2005 ◽  
Vol 33 (4) ◽  
pp. 802-805 ◽  
Author(s):  
S. Mann ◽  
A. Marquet ◽  
O. Ploux
Keyword(s):  
Active Site ◽  
Drug Target ◽  
1H Nmr Spectroscopy ◽  
Biochemical Characterization ◽  
Membered Ring ◽  
Potential Drug Target ◽  
Visible Spectroscopy ◽  
X Ray ◽  
Uv Visible ◽  
Cis And Trans

Cis and trans stereoisomers of amiclenomycin, a natural L-amino acid antibiotic, have been prepared using unequivocal routes. By using 1H NMR spectroscopy, the configuration of the six-membered ring of natural amiclenomycin was shown to be cis and not trans as originally proposed. Amiclenomycin and some synthetic analogues with the cis configuration irreversibly inactivate DAPA AT (7,8-diaminopelargonic acid aminotransferase), an enzyme involved in biotin biosynthesis, by forming an aromatic PLP (pyridoxal-5′-phosphate)–inhibitor adduct that is tightly bound to the active site. The following kinetic parameters for the inactivation of Escherichia coli DAPA AT by amiclenomycin were derived: KI=2 μM and kinact=0.4 min−1. The structure of the aromatic adduct formed upon inactivation was confirmed by UV–visible spectroscopy, X-ray crystal structure determination and MS. Because Mycobacterium tuberculosis DAPA AT is a potential drug target, this enzyme was cloned, overexpressed and purified to homogeneity for biochemical characterization.

scholarly journals Structural Basis for Inhibition of Cathepsin B Drug Target from the Human Blood Fluke, Schistosoma mansoni

2011 ◽  
Vol 286 (41) ◽  
pp. 35770-35781 ◽  
Author(s):  
Adéla Jílková ◽  
Pavlína Řezáčová ◽  
Martin Lepšík ◽  
Martin Horn ◽  
Jana Váchová ◽  
...  
Keyword(s):  
Schistosoma Mansoni ◽  
Active Site ◽  
Drug Target ◽  
Rational Design ◽  
Structural Basis ◽  
Vinyl Sulfone ◽  
Blood Proteins ◽  
Blood Fluke ◽  
Active Site Cleft ◽  
Host Blood

Schistosomiasis caused by a parasitic blood fluke of the genus Schistosoma afflicts over 200 million people worldwide. Schistosoma mansoni cathepsin B1 (SmCB1) is a gut-associated peptidase that digests host blood proteins as a source of nutrients. It is under investigation as a drug target. To further this goal, we report three crystal structures of SmCB1 complexed with peptidomimetic inhibitors as follows: the epoxide CA074 at 1.3 Å resolution and the vinyl sulfones K11017 and K11777 at 1.8 and 2.5 Å resolutions, respectively. Interactions of the inhibitors with the subsites of the active-site cleft were evaluated by quantum chemical calculations. These data and inhibition profiling with a panel of vinyl sulfone derivatives identify key binding interactions and provide insight into the specificity of SmCB1 inhibition. Furthermore, hydrolysis profiling of SmCB1 using synthetic peptides and the natural substrate hemoglobin revealed that carboxydipeptidase activity predominates over endopeptidolysis, thereby demonstrating the contribution of the occluding loop that restricts access to the active-site cleft. Critically, the severity of phenotypes induced in the parasite by vinyl sulfone inhibitors correlated with enzyme inhibition, providing support that SmCB1 is a valuable drug target. The present structure and inhibitor interaction data provide a footing for the rational design of anti-schistosomal inhibitors.

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