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JournalISSN: 1554-8929

ACS Chemical Biology 

American Chemical Society
About: ACS Chemical Biology is an academic journal published by American Chemical Society. The journal publishes majorly in the area(s): Medicine & RNA. It has an ISSN identifier of 1554-8929. Over the lifetime, 4452 publications have been published receiving 158910 citations. The journal is also known as: ACS Chem. Biol. & ACS Chem Biol.
Topics: Medicine, RNA, Chemistry, Peptide, Small molecule


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Journal ArticleDOI
TL;DR: The utility of this modular protein tagging system for cellular imaging and protein immobilization is demonstrated by analyzing multiple molecular processes associated with NF-kappaB-mediated cellular physiology, including imaging of subcellular protein translocation and capture of protein--protein and protein--DNA complexes.
Abstract: We have designed a modular protein tagging system that allows different functionalities to be linked onto a single genetic fusion, either in solution, in living cells, or in chemically fixed cells. The protein tag (HaloTag) is a modified haloalkane dehalogenase designed to covalently bind to synthetic ligands (HaloTag ligands). The synthetic ligands comprise a chloroalkane linker attached to a variety of useful molecules, such as fluorescent dyes, affinity handles, or solid surfaces. Covalent bond formation between the protein tag and the chloroalkane linker is highly specific, occurs rapidly under physiological conditions, and is essentially irreversible. We demonstrate the utility of this system for cellular imaging and protein immobilization by analyzing multiple molecular processes associated with NF-κB-mediated cellular physiology, including imaging of subcellular protein translocation and capture of protein−protein and protein−DNA complexes.

1,822 citations

Journal ArticleDOI
TL;DR: A novel bioluminescence system capable of more efficient light emission with superior biochemical and physical characteristics is engineered in mammalian cells by merging optimization of protein structure with development of a novel imidazopyrazinone substrate (furimazine).
Abstract: Bioluminescence methodologies have been extraordinarily useful due to their high sensitivity, broad dynamic range, and operational simplicity. These capabilities have been realized largely through incremental adaptations of native enzymes and substrates, originating from luminous organisms of diverse evolutionary lineages. We engineered both an enzyme and substrate in combination to create a novel bioluminescence system capable of more efficient light emission with superior biochemical and physical characteristics. Using a small luciferase subunit (19 kDa) from the deep sea shrimp Oplophorus gracilirostris, we have improved luminescence expression in mammalian cells ∼2.5 million-fold by merging optimization of protein structure with development of a novel imidazopyrazinone substrate (furimazine). The new luciferase, NanoLuc, produces glow-type luminescence (signal half-life >2 h) with a specific activity ∼150-fold greater than that of either firefly (Photinus pyralis) or Renilla luciferases similarly conf...

1,166 citations

Journal ArticleDOI
TL;DR: The chemical and photophysical properties of oft-used fluorophores are examined and classic and contemporary examples in which utility has been built upon these scaffolds are highlighted.
Abstract: Small-molecule fluorescent probes embody an essential facet of chemical biology. Although numerous compounds are known, the ensemble of fluorescent probes is based on a modest collection of modular “core” dyes. The elaboration of these dyes with diverse chemical moieties is enabling the precise interrogation of biochemical and biological systems. The importance of fluorescence-based technologies in chemical biology elicits a necessity to understand the major classes of small-molecule fluorophores. Here, we examine the chemical and photophysical properties of oft-used fluorophores and highlight classic and contemporary examples in which utility has been built upon these scaffolds.

1,086 citations

Journal ArticleDOI
TL;DR: NanoBiT provided a means to measure pharmacology of kinase inhibitors known to induce the interaction between BRAF and CRAF and that the reporter responds reliably and dynamically in cells.
Abstract: Protein-fragment complementation assays (PCAs) are widely used for investigating protein interactions. However, the fragments used are structurally compromised and have not been optimized nor thoroughly characterized for accurately assessing these interactions. We took advantage of the small size and bright luminescence of NanoLuc to engineer a new complementation reporter (NanoBiT). By design, the NanoBiT subunits (i.e., 1.3 kDa peptide, 18 kDa polypeptide) weakly associate so that their assembly into a luminescent complex is dictated by the interaction characteristics of the target proteins onto which they are appended. To ascertain their general suitability for measuring interaction affinities and kinetics, we determined that their intrinsic affinity (KD = 190 μM) and association constants (kon = 500 M–1 s–1, koff = 0.2 s–1) are outside of the ranges typical for protein interactions. The accuracy of NanoBiT was verified under defined biochemical conditions using the previously characterized interaction...

817 citations

Journal ArticleDOI
TL;DR: An "interfacial activity model" is proposed, which is based on an experimentally testable molecular image of AMP-membrane interactions, which may be useful in driving engineering and design of novel AMPs.
Abstract: Antimicrobial peptides (AMPs) have been studied for three decades, and yet a molecular understanding of their mechanism of action is still lacking. Here we summarize current knowledge for both synthetic vesicle experiments and microbe experiments, with a focus on comparisons between the two. Microbial experiments are done at peptide to lipid ratios that are at least 4 orders of magnitude higher than vesicle-based experiments. To close the gap between the two concentration regimes, we propose an “interfacial activity model”, which is based on an experimentally testable molecular image of AMP–membrane interactions. The interfacial activity model may be useful in driving engineering and design of novel AMPs.

788 citations

Performance
Metrics
No. of papers from the Journal in previous years
YearPapers
2023176
2022381
2021281
2020346
2019311
2018375