Abstract：
:A biophysical understanding of the mechanistic, chemical, and physical origins underlying antibiotic action and resistance is vital to the discovery of novel therapeutics and the development of strategies to combat the growing emergence of antibiotic resistance. The site-specific introduction of stable-isotope labels into chemically complex natural products is particularly important for techniques such as NMR, IR, mass spectrometry, imaging, and kinetic isotope effects. Toward this goal, we developed a biosynthetic strategy for the site-specific incorporation of 13C labels into the canonical β-lactam carbonyl of penicillin G and cefotaxime, the latter via cephalosporin C. This was achieved through sulfur-replacement with 1-13C-l-cysteine, resulting in high isotope incorporations and milligram-scale yields. Using 13C NMR and isotope-edited IR difference spectroscopy, we illustrate how these molecules can be used to interrogate interactions with their protein targets, e.g., TEM-1 β-lactamase. This method provides a feasible route to isotopically labeled penicillin and cephalosporin precursors for future biophysical studies.
journal_name
ACS Chem Bioljournal_title
ACS chemical biologyauthors
Kozuch J,Schneider SH,Boxer SGdoi
10.1021/acschembio.9b01054subject
Has Abstractpub_date
2020-05-15 00:00:00pages
1148-1153issue
5eissn
1554-8929issn
1554-8937journal_volume
15pub_type
杂志文章abstract：:The Morita-Baylis-Hillman reaction forms a carbon-carbon bond between the α-carbon of a conjugated carbonyl compound and a carbon electrophile. The reaction mechanism involves Michael addition of a nucleophile catalyst at the carbonyl β-carbon, followed by bond formation with the electrophile and catalyst disassociati...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/cb3006227
更新日期：2013-04-19 00:00:00
abstract：:Beta-peptides (beta-amino acid oligomers) that mimic the amphiphilic, helical, and cationic properties of natural antimicrobial peptides have previously been shown to display antifungal activity against planktonic Candida albicans cells. Beta-peptides offer several advantages over conventional peptides composed of alp...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/cb900093r
更新日期：2009-07-17 00:00:00
abstract：:A complication of diabetes is the inability of wounds to heal in diabetic patients. Diabetic wounds are refractory to healing due to the involvement of activated matrix metalloproteinases (MMPs), which remodel the tissue resulting in apoptosis. There are no readily available methods that identify active unregulated MM...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/cb4005468
更新日期：2014-01-17 00:00:00
abstract：:New machine learning methods to analyze raw chemical and biological data are now widely accessible as open-source toolkits. This positions researchers to leverage powerful, predictive models in their own domains. We caution, however, that the application of machine learning to experimental research merits careful cons...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/acschembio.8b00881
更新日期：2018-10-19 00:00:00
abstract：:mRNA-based therapies and vaccines constitute a disruptive technology with the potential to revolutionize modern medicine. Chemically modified 5' cap structures have provided access to mRNAs with superior translational properties that could benefit the currently flourishing mRNA field. Prime examples of compounds that ...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/acschembio.0c00864
更新日期：2021-01-13 00:00:00
abstract：:The nematode Caenorhabditis elegans is an excellent model organism for studies of glycan dynamics, a goal that requires tools for imaging glycans in vivo. Here we applied the bioorthogonal chemical reporter technique for the molecular imaging of mucin-type O-glycans in live C. elegans. We treated worms with azidosugar...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/cb900254y
更新日期：2009-12-18 00:00:00
abstract：:Gaussia luciferase (GLUC) is a bioluminescent reporter protein of increasing importance. As a secretory protein, it has increased sensitivity in vitro and in vivo (∼20 000-fold, and ∼1000-fold, respectively) over its competitor, secreted alkaline phosphatase. Unfortunately, this same advantageous secretory nature of G...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/acschembio.7b00454
更新日期：2017-09-15 00:00:00
abstract：:Dihydrofolate reductase (DHFR) is a potential drug target for Trypanosoma brucei, a human parasite, which is the causative agent for African sleeping sickness. No drug is available against this target, since none of the classical antifolates such as pyrimethamine (PYR), cycloguanil, or trimethoprim are effective as se...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/cb200124r
更新日期：2011-09-16 00:00:00
abstract：:Enzymatic plasticity, as a modern term referring to the functional conversion of an enzyme, is significant for enzymatic activity redesign. The bacterial diterpene cyclase CotB2 is a typical plastic enzyme by which its native form precisely conducts a chemical reaction while its mutants diversify the catalytic functio...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/acschembio.0c00645
更新日期：2020-10-16 00:00:00
abstract：:Nonribosomal peptides are important natural products biosynthesized by nonribosomal peptide synthetases (NRPSs). Adenylation (A) domains of NRPSs are highly specific for the substrate they recognize. This recognition is determined by 10 residues in the substrate-binding pocket, termed the specificity code. This findin...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/acschembio.9b00532
更新日期：2019-09-20 00:00:00
abstract：:How primitive enzymes emerged from a primordial pool remains a fundamental unanswered question with important practical implications in synthetic biology. Here we show that a de novo evolved ATP binding protein, selected solely on the basis of its ability to bind ATP, mediates the regiospecific hydrolysis of ATP to AD...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/cb900109w
更新日期：2009-08-21 00:00:00
abstract：:Toxic heavy metals have been considered to be harmful environmental contaminations. The molecular mechanisms of heavy-metals-induced cytotoxicity and carcinogenicity are still not well elucidated. Previous reports showed exposures to toxic heavy metals can cause a change of DNA cytosine methylation (5-methylcytosine, ...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/acschembio.7b00170
更新日期：2017-06-16 00:00:00
abstract：:Hypoxia induces a complex circuit of gene expression that drives tumor progression and increases drug resistance. Defining these changes allows for an understanding of how hypoxia alters tumor biology and informs design of lead therapeutics. We probed the role of microRNA-544 (miR-544), which silences mammalian target...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/acschembio.5b00265
更新日期：2015-10-16 00:00:00
abstract：:The genetic code specifies 20 common amino acids and is largely preserved in both single and multicellular organisms. Unnatural amino acids (Uaas) have been genetically incorporated into proteins by using engineered orthogonal tRNA/aminoacyl-tRNA synthetase (RS) pairs, enabling new research capabilities and precision ...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/cb200542j
更新日期：2012-07-20 00:00:00
abstract：:Regulation of gene expression by RNA processing mechanisms is now understood to be an important level of control in mammalian cells. Regulation at the level of RNA transcription, splicing, polyadenylation, nucleo-cytoplasmic transport, and translation into polypeptides has been well-studied. Alternative RNA processing...
journal_title：ACS chemical biology
pub_type： 杂志文章,评审
doi：10.1021/cb800138w
更新日期：2008-10-17 00:00:00
abstract：:The flavo-enzyme DprE1 catalyzes a key epimerization step in the decaprenyl-phosphoryl d-arabinose (DPA) pathway, which is essential for mycobacterial cell wall biogenesis and targeted by several new tuberculosis drug candidates. Here, using differential radiolabeling with DPA precursors and high-resolution fluorescen...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/acschembio.5b00237
更新日期：2015-07-17 00:00:00
abstract：:Diversity of scaffold structure and function is a hallmark of the >50,000 isoprenoid natural products such as taxol. Whereas most members of this class are assembled by iterative head-to-tail enzymatic joining reactions between delta2- and delta3-isopentenyl diphosphate (IPP) monomers, dimerization of two delta2-IPP m...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/cb700094s
更新日期：2007-05-22 00:00:00
abstract：:Transcriptomes provide a myriad of potential RNAs that could be the targets of therapeutics or chemical genetic probes of function. Cell-permeable small molecules, however, generally do not exploit these targets, owing to the difficulty in the design of high affinity, specific small molecules targeting RNA. As part of...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/cb400265y
更新日期：2013-10-18 00:00:00
abstract：:Phosphatases play key roles in normal physiology and diseases. Studying phosphatases has been both essential and challenging, and the application of conventional genetic and biochemical methods has led to crucial but still limited understanding of their mechanisms, substrates, and exclusive functions within highly int...
journal_title：ACS chemical biology
pub_type： 杂志文章,评审
doi：10.1021/acschembio.6b00570
更新日期：2016-11-18 00:00:00
abstract：:The perturbation of protein kinases with small organic molecules is a powerful approach to dissect kinase function in complex biological systems. Covalent kinase inhibitors that target thiols in the ATP binding pocket of the kinase domain proved to be ideal reagents for the investigation of highly dynamic cellular pro...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/cb200465c
更新日期：2012-04-20 00:00:00
abstract：:Recent advances in mass spectrometry (MS)-based proteomics allow the identification and quantitation of thousands of posttranslational modification (PTM) sites in a single experiment. This follows from the development of more effective class enrichment strategies, new high performance instrumentation and bioinformatic...
journal_title：ACS chemical biology
pub_type： 杂志文章,评审
doi：10.1021/cb500904b
更新日期：2015-01-16 00:00:00
abstract：:Bacteria use small molecules to assess the density and identity of nearby organisms and formulate a response. This process, called quorum sensing (QS), commonly regulates bioluminescence, biofilm formation, and virulence. Vibrio harveyi have three described QS circuits. Each involves the synthesis of a molecule that r...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/cb300215t
更新日期：2012-08-17 00:00:00
abstract：:Huntington's disease (HD) is a neurodegenerative disorder that is caused by abnormal expansion of CAG repeats in the HTT gene. The transcribed mutant RNA contains expanded CAG repeats that translate into a mutant huntingtin protein. This expanded CAG repeat also causes mis-splicing of pre-mRNA due to sequestration of ...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/acschembio.7b00699
更新日期：2018-01-19 00:00:00
abstract：:Alzheimer's Disease (AD) is a progressive neurodegenerative disease and the most common cause of dementia. The current treatment options for AD are limited to ameliorating cognitive decline temporarily and not reversing or preventing the progression of dementia. Hence, more effective therapeutic strategies are needed ...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/acschembio.0c00851
更新日期：2020-12-18 00:00:00
abstract：:In eukaryotic cells, proteins and RNAs are transported between the nucleus and the cytoplasm by nuclear import and export receptors. Over the past decade, small molecules that inhibit the nuclear export receptor CRM1 have been identified, most notably leptomycin B. However, up to now no small molecule inhibitors of nu...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/cb100094k
更新日期：2010-10-15 00:00:00
abstract：:The innate immune system provides the first line of defense against pathogens through the recognition of nonspecific patterns in RNA to protect the cell in a generalized way. The human RNA-activated protein kinase, PKR, is a dsRNA binding protein and an essential sensor in the innate immune response, which recognizes ...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/acschembio.6b00081
更新日期：2016-04-15 00:00:00
abstract：:The heterologous biosynthesis of complex natural products has enabled access to polyketide, nonribosomal peptide, isoprenoid, and other compounds with wide-spanning societal value. Though several surrogate host systems exist, Escherichia coli is often a preferred choice due to its rapid growth kinetics and extensive m...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/acschembio.9b00827
更新日期：2020-05-15 00:00:00
abstract：:FAT10 is a ubiquitin-like protein suggested to target proteins for proteasomal degradation. It is highly upregulated upon pro-inflammatory cytokines, namely, TNFα, IFNγ, and IL6, and was found to be highly expressed in various epithelial cancers. Evidence suggests that FAT10 is involved in cancer development and may h...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/acschembio.9b00667
更新日期：2019-12-20 00:00:00
abstract：:Determining the impact of lipid electrophile-mediated protein damage that occurs during oxidative stress requires a comprehensive analysis of electrophile targets adducted under pathophysiological conditions. Incorporation of ω-alkynyl linoleic acid into the phospholipids of macrophages prior to activation by Kdo2-lip...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/acschembio.7b00480
更新日期：2017-08-18 00:00:00
abstract：:Siderophores are iron-chelating molecules produced by microorganisms and plants to acquire exogenous iron. Siderophore biosynthetic enzymology often produces elaborate and unique molecules through unusual reactions to enable specific recognition by the producing organisms. Herein, we report the structure of two sidero...
journal_title：ACS chemical biology
pub_type： 杂志文章
doi：10.1021/acschembio.0c00809
更新日期：2021-01-15 00:00:00
