112921-00-3Relevant articles and documents
An orthogonal seryl-tRNA synthetase/tRNA pair for noncanonical amino acid mutagenesis in Escherichia coli
Zambaldo, Claudio,Koh, Minseob,Nasertorabi, Fariborz,Han, Gye Won,Chatterjee, Abhishek,Stevens, Raymond C.,Schultz, Peter G.
, (2020)
We report the development of the orthogonal amber-suppressor pair Archaeoglobus fulgidus seryl-tRNA (Af-tRNASer)/Methanosarcina mazei seryl-tRNA synthetase (MmSerRS) in Escherichia coli. Furthermore, the crystal structure of MmSerRS was solved at 1.45 ? resolution, which should enable structure-guided engineering of its active site to genetically encode small, polar noncanonical amino acids (ncAAs).
Discovery of Leucyladenylate Sulfamates as Novel Leucyl-tRNA Synthetase (LRS)-Targeted Mammalian Target of Rapamycin Complex 1 (mTORC1) Inhibitors
Yoon, Suyoung,Kim, Jong Hyun,Kim, Sung-Eun,Kim, Changhoon,Tran, Phuong-Thao,Ann, Jihyae,Koh, Yura,Jang, Jayun,Kim, Sungmin,Moon, Hee-Sun,Kim, Won Kyung,Lee, Sangkook,Lee, Jiyoun,Kim, Sunghoon,Lee, Jeewoo
, p. 10322 - 10328 (2016)
Recent studies indicate that LRS may act as a leucine sensor for the mTORC1 pathway, potentially providing an alternative strategy to overcome rapamycin resistance in cancer treatments. In this study, we developed leucyladenylate sulfamate derivatives as LRS-targeted mTORC1 inhibitors. Compound 18 selectively inhibited LRS-mediated mTORC1 activation and exerted specific cytotoxicity against colon cancer cells with a hyperactive mTORC1, suggesting that 18 may offer a novel treatment option for human colorectal cancer.
A FACILE SYNTHESYS OF ASCAMYCIN AND RELATED ANALOGUES
Castro-Pichel, Julia,Garcia-Lopez, Maria Teresa,De las Heras, Federico G.
, p. 383 - 390 (1987)
The nucleoside antibiotic ascamycin (2-chloro-5'-O-(N-(L-alanyl)sulfamoyl)adenosine (1)) has been synthesized by an improved procedure involving the direct condensation of 2-chloro-2',3'-O-isopropylidene-5'-O-sulfamoyladenosine (3) with Boc-L-Ala-OSu in DMF and the presence of DBU, followed by removal of the protecting groups.A similar condensation of 3 with Boc-D-Ala-Osu and Boc-Gly-Osu, and subsequent deprotection, yielded the D-Ala and Gly analogues of 1, namely 2-chloro-5'-O-(N-(D-alanyl)sulfamoyl) and 2-chloro-5'-O-(N-(glycyl)sulfamoyl)adenosine (D-ascamycin (14) and (18)).Similar reactions of 2',3'-O-isopropylidene-5'-O-sulfamoyladenosine, (6) with the tree amino acid derivatives above mentioned provided the corresponding adenosine analogues 12,16, and 20.Several studies directed to demonstrate that the previous protection of the 6-NH2 group of the adenosine derivatives 3 and 6 is not necessary for the selective aminoacylation of the SO2NH2 group are also reported.
Synthesis, structure and bioactivity of primary sulfamate-containing natural products
Mujumdar, Prashant,Bua, Silvia,Supuran, Claudiu T.,Peat, Thomas S.,Poulsen, Sally-Ann
, p. 3009 - 3013 (2018)
Here we report the synthesis of natural products (NPs) 5′-O-sulfamoyl adenosine 1 and 5′-O-sulfamoyl-2-chloroadenosine 2. As primary sulfamates these compounds represent an uncommon class of NPs, furthermore there are few NPs known that contain a N–S bond. Compounds 1 and 2 were evaluated for inhibition of carbonic anhydrases (CA), a metalloenzyme family where the primary sulfamate is known to coordinate to the active site zinc and form key hydrogen bonds with adjacent CA active site residues. Both NPs were good to moderate CA inhibitors, with compound 2 a 20–50-fold stronger CA inhibitor (Ki values 65–234 nM) than compound 1. The protein X-ray crystal structures of 1 and 2 in complex with CA II show that it is not the halogen-hydrophobic interactions that give compound 2 a greater binding energy but a slight movement in orientation of the ribose ring that allows better hydrogen bonds to CA residues. Compounds 1 and 2 were further investigated for antimicrobial activity against a panel of microbes relevant to human health, including Gram-negative bacteria (4 strains), Gram-positive bacteria (1 strain) and yeast (2 strains). Antimicrobial activity and selectivity was observed. The minimum inhibitory concentration (MIC) of NP 1 was 10 μM against Gram-positive Staphylococcus aureus and NP 2 was 5 μM against Gram-negative Escherichia coli. This is the first time that NP primary sulfamates have been assessed for inhibition and binding to CAs, with systematic antimicrobial activity studies also reported.
Synthesis and aminoacyl-tRNA synthetase inhibitory activity of prolyl adenylate analogs
Heacock, Donald,Forsyth, Craig J.,Shiba, Kiyotaka,Musier-Forsyth, Karin
, p. 273 - 289 (1996)
Two nonhydrolyzable prolyl adenylate analogs, 5'-O-[N-(L-prolyl)-sulfamoyl]adenosine (L-PSA) and 5'-O-[N-(D-prolyl)-sulfamoyl]adenosine (D-PSA), were prepared in three steps from 2',3'-di-O-isopropylideneadenosine. Both of these compounds inhibited the in vitro activity of Escherichia coli and human prolyl-tRNA synthetase (ProRS). The human enzyme used in this study was derived from the carboxy-terminal domain of the multifunctional human EPRS gene. The K(i)(ATP) values for L-PSA, determined using the ATP-PP(i) exchange assay, are very similar for both synthetases (~1-2 nM). The K(i)(Pro) values, on the other hand, vary approximately seven-fold between the two synthetases (0.6 nM for human and 4.3 nM for E. coli). The K(i) values measured for the D-PSA analog are much higher (51-470 nM) for all cases examined; however, the same species-specific differences are observed with respect to K(i)(Pro). These results indicate possible structural differences in or near the active sites of the two enzymes that may be exploited in the future design of compounds that function as species-specific synthetase inhibitors in vivo.
Dissecting modular synthases through inhibition: A complementary chemical and genetic approach
Vickery, Christopher R.,McCulloch, Ian P.,Sonnenschein, Eva C.,Beld, Joris,Noel, Joseph P.,Burkart, Michael D.
, (2019/12/24)
Modular synthases, such as fatty acid, polyketide, and non-ribosomal peptide synthases (NRPSs), are sophisticated machineries essential in both primary and secondary metabolism. Various techniques have been developed to understand their genetic background and enzymatic abilities. However, uncovering the actual biosynthetic pathways remains challenging. Herein, we demonstrate a pipeline to study an assembly line synthase by interrogating the enzymatic function of each individual enzymatic domain of BpsA, a NRPS that produces the blue 3,3′-bipyridyl pigment indigoidine. Specific inhibitors for each biosynthetic domain of BpsA were obtained or synthesized, and the enzymatic performance of BpsA upon addition of each inhibitor was monitored by pigment development in vitro and in living bacteria. The results were verified using genetic mutants to inactivate each domain. Finally, the results complemented the currently proposed biosynthetic pathway of BpsA.
Integrated Target-Based and Phenotypic Screening Approaches for the Identification of Anti-Tubercular Agents That Bind to the Mycobacterial Adenylating Enzyme MbtA
Ferguson, Lindsay,Wells, Geoff,Bhakta, Sanjib,Johnson, James,Guzman, Junitta,Parish, Tanya,Prentice, Robin A.,Brucoli, Federico
supporting information, p. 1735 - 1741 (2019/10/22)
Iron is essential for the pathogenicity and virulence of Mycobacterium tuberculosis, which synthesises salicyl-capped siderophores (mycobactins) to acquire this element from the host. MbtA is the adenylating enzyme that catalyses the initial reaction of mycobactin biosynthesis and is solely expressed by mycobacteria. A 3200-member library comprised of lead-like, structurally diverse compounds was screened against M. tuberculosis for whole-cell inhibitory activity. A set of 846 compounds that inhibited the tubercle bacilli growth were then tested for their ability to bind to MbtA using a fluorescence-based thermal shift assay and NMR-based Water-LOGSY and saturation transfer difference (STD) experiments. We identified an attractive hit molecule, 5-hydroxyindol-3-ethylamino-(2-nitro-4-trifluoromethyl)benzene (5), that bound with high affinity to MbtA and produced a MIC90 value of 13 μm. The ligand was docked into the MbtA crystal structure and displayed an excellent fit within the MbtA active pocket, adopting a binding mode different from that of the established MbtA inhibitor Sal-AMS.
Structure-activity relationship of adenosine 5′-diphosphoribose at the transient receptor potential melastatin 2 (TRPM2) channel: Rational design of antagonists
Moreau, Christelle,Kirchberger, Tanja,Swarbrick, Joanna M.,Bartlett, Stephen J.,Fliegert, Ralf,Yorgan, Timur,Bauche, Andreas,Harneit, Angelika,Guse, Andreas H.,Potter, Barry V. L.
supporting information, p. 10079 - 10102 (2014/01/17)
Adenosine 5′-diphosphoribose (ADPR) activates TRPM2, a Ca 2+, Na+, and K+ permeable cation channel. Activation is induced by ADPR binding to the cytosolic C-terminal NudT9-homology domain. To generate the first structure-activity relationship, systematically modified ADPR analogues were designed, synthesized, and evaluated as antagonists using patch-clamp experiments in HEK293 cells overexpressing human TRPM2. Compounds with a purine C8 substituent show antagonist activity, and an 8-phenyl substitution (8-Ph-ADPR, 5) is very effective. Modification of the terminal ribose results in a weak antagonist, whereas its removal abolishes activity. An antagonist based upon a hybrid structure, 8-phenyl-2′-deoxy-ADPR (86, IC50 = 3 μM), is more potent than 8-Ph-ADPR (5). Initial bioisosteric replacement of the pyrophosphate linkage abolishes activity, but replacement of the pyrophosphate and the terminal ribose by a sulfamate-based group leads to a weak antagonist, a lead to more drug-like analogues. 8-Ph-ADPR (5) inhibits Ca2+ signalling and chemotaxis in human neutrophils, illustrating the potential for pharmacological intervention at TRPM2.
Enantiodifferentiation of ketoprofen by Japanese firefly luciferase from Luciola lateralis
Kato, Dai-Ichiro,Tatsumi, Tomohiro,Bansho, Asami,Teruya, Keisuke,Yoshida, Hiromitsu,Takeo, Masahiro,Negoro, Seiji
experimental part, p. 140 - 146 (2012/01/19)
Recently, we found that firefly luciferase exhibited (R)-enantioselective thioesterification activity toward 2-arylpropanoic acids. In the case of Japanese firefly luciferase from Luciola lateralis (LUC-H), the E-value for ketoprofen was approximately 20. In this study, we used a spectrophotometric method to measure the catalytic activity of LUC-H. Using this method allowed us to judge the reaction efficiency easily. Our results confirmed that LUC-H exhibits enantioselective thioesterification activity toward a series of 2-arylpropanoic acids. The highest activity was observed with ketoprofen. We also observed high enzymatic activity of LUC-H toward long-chain fatty acids. These results were reasonable because LUC-H is homologous with long-chain acyl-CoA synthetase. To obtain further information about the enantiodifferentiation mechanism of the LUC-H catalyzed thioesterification of ketoprofen, we determined the kinetic parameters of the reaction relative to each of its three substrates: ketoprofen, ATP, and coenzyme A (CoASH). We found that whereas the affinities of each compound are not affected by the chirality of ketoprofen, enantiodifferentiation is achieved by a chirality-dependent difference in the kcat parameter.
Identification of NAE inhibitors exhibiting potent activity in leukemia cells: Exploring the structural determinants of NAE specificity
Lukkarila, Julie L.,Da Silva, Sara R.,Ali, Mohsin,Shahani, Vijay M.,Xu, G. Wei,Berman, Judd,Roughton, Andrew,Dhe-Paganon, Sirano,Schimmer, Aaron D.,Gunning, Patrick T.
supporting information; experimental part, p. 577 - 582 (2011/10/02)
MLN4924 is a selective inhibitor of the NEDD8-activating enzyme (NAE) and has advanced into clinical trials for the treatment of both solid and hematological malignancies. In contrast, the structurally similar compound 1 (developed by Millennium: The Takeda Oncology Company) is a pan inhibitor of the E1 enzymes NAE, ubiquitin activating enzyme (UAE), and SUMO-activating enzyme (SAE) and is currently viewed as unsuitable for clinical use given its broad spectrum of E1 inhibition. Here, we sought to understand the determinants of NAE selectivity. A series of compound 1 analogues were synthesized through iterative functionalization of the purine C6 position and evaluated for NAE specificity. Optimal NAE specificity was achieved through substitution with primary N-alkyl groups, while bulky or secondary N-alkyl substituents were poorly tolerated. When assessed in vitro, inhibitors reduced the growth and viability of malignant K562 leukemia cells. Through this study, we have successfully identified a series of sub-10 nM NAE-specific inhibitors and thereby highlighted the functionalities that promote NAE selectivity.
