A Direct Method for Site‐Specific Protein Acetylation

Li, Fupeng; Allahverdi, Abdollah; Yang, Renliang; Lua, Gavian Bing Jia; Zhang, Xiaohong; Cao, Yuan; Korolev, Nikolay; Nordenskiöld, Lars; Liu, Chuan‐Fa

doi:10.1002/anie.201103754

Cited by 128 publications

(103 citation statements)

References 40 publications

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“…44,45 The linker connecting the dextran and thiol in D1 is located at the reducing end of the dextran and contains amide and ether moieties. All concentrations of conjugate 2 were measured as total dextran concentration (labeled and unlabeled), and all three batches showed identical same staining patterns and qualitative results.…”

Section: Methodsmentioning

confidence: 99%

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Cytosolic Uptake of Large Monofunctionalized Dextrans

2018

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Dextrans are a versatile class of polysaccharides with applications that span medicine, cell biology, food science, and consumer goods. Here, we report on a new type of large monofunctionalized dextran that exhibits unusual properties: efficient cytosolic and nuclear uptake. This dextran permeates various human cell types without the use of transfection agents, electroporation, or membrane perturbation. Cellular uptake occurs primarily through active transport via receptor-mediated processes. These monofunctionalized dextrans could serve as intracellular delivery platforms for drugs or other cargos.

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Section: Methodsmentioning

confidence: 99%

“…Glutathione (1.5 equiv), lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP) initiator 44,45 (2.5 equiv), and probe 1a (5 equiv) were added. The reaction mixture was irradiated with a 365-nm light source for 15 min.…”

Section: Methodsmentioning

confidence: 99%

Cytosolic Uptake of Large Monofunctionalized Dextrans

2018

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“…18 These include reaction with 1) monobromomaleimide, 27 2) vinyl sulfones 28 and 3) allenamides, 29 or 4) somewhat conversely modification to dehydroalanine followed by Michael addition reaction with thiol. 30 Other thiol-selective reactions include 5) light initiated thiol-ene, 31 6) perfluoroaromatic molecules 32 and 7) sulfone reagents that are Julia-Kocienski-like 33 . Using these thiol modification reactions in ADC synthesis may improve the species homogeneity.…”

Section: Conjugation Methodsmentioning

confidence: 99%

Field Guide to Challenges and Opportunities in Antibody–Drug Conjugates for Chemists

Gordon

Canakci

et al. 2015

Bioconjugate Chem.

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Antibody-drug conjugates have attracted a great amount of attention as a therapeutic strategy for diseases where targeting specific tissues and cells are critical components, such as in cancer therapy. Although promising, the number of approved ADC drugs is relatively limited. This emanates from the challenges associated with generating the conjugates and the complexities associated with the stability requirements for these conjugates during circulation and after reaching the target. Here, we provide a comprehensive overview of the design challenges facing the ADC field. These challenges also provide several unique research and development opportunities, which are also highlighted throughout the review.

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“…[5] However,w hile most of these transformations deliver excellent results on peptides and recombinant proteins,o nly very few of them have been shown to be also effective in labeling cysteines in cells or cellular lysates and therefore suitable for proteomics applications.I odoacetamide (IAA) is widely accepted as gold standard among MS-compatible cysteinereactive chemical probes, [6] but it displays rather low chemical stability in aqueous buffers and it only reacts with afraction of functional cysteines. [7] Moreover,i odoacetamide also modifies lysines and the formed artifacts lead to misassignment of lysine ubiquitination sites by mass spectrometry.…”

Section: Despitetheirrelativelylowabundanceinproteinscysteinesmentioning

confidence: 99%

Proteome‐Wide Profiling of Targets of Cysteine reactive Small Molecules by Using Ethynyl Benziodoxolone Reagents

et al. 2015

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In this study,wepresent ahighly efficient method for proteomic profiling of cysteine residues in complex proteomes and in living cells.O ur method is based on alkynylation of cysteines in complex proteomes using a" clickable" alkynyl benziodoxolone bearing an azide group.T his reaction proceeds fast, under mild physiological conditions,and with avery high degree of chemoselectivity.T he formed azide-capped alkynyl-cysteine adducts are readily detectable by LC-MS/MS, and can be further functionalizedw ith TAMRA or biotin alkyne via CuAAC. We demonstrate the utility of alkynyl benziodoxolones for chemical proteomics applications by identifying the proteomic targets of curcumin, ad iarylheptanoid natural product that was and still is part of multiple human clinical trials as anticancer agent. Our results demonstrate that curcumin covalently modifies several key players of cellular signaling and metabolism, most notably the enzyme casein kinase Igamma. We anticipate that this new method for cysteine profiling will find broad application in chemical proteomics and drug discovery. Despitetheirrelativelylowabundanceinproteins,cysteinesare vital for cellular biochemistry.F or example,c ysteines form disulfide bridges,a re actively involved in enzyme catalysis as nucleophiles and can be posttranslationally oxidized or modified through palmitoylation, prenylation or nitrosylation.[1] Many of the so-called functional cysteines display pronounced hyperreactivity and these reactive hot spots can be precisely identified in complex proteomes with broad-range electrophilic probes.[2] Such probes represent particularly useful tools for competitive activity-based protein profiling [3] (ABPP) of targets of covalently binding cysteinereactive drugs and metabolites,asimpressively demonstrated in recent publications.[4] Over the past few years,abroad variety of novel methods for broad-spectrum, yet chemoselective targeting of cysteiner esidues have been reported. [5] However,w hile most of these transformations deliver excellent results on peptides and recombinant proteins,o nly very few of them have been shown to be also effective in labeling cysteines in cells or cellular lysates and therefore suitable for proteomics applications.I odoacetamide (IAA) is widely accepted as gold standard among MS-compatible cysteinereactive chemical probes, [6] but it displays rather low chemical stability in aqueous buffers and it only reacts with afraction of functional cysteines. [7] Moreover,i odoacetamide also modifies lysines and the formed artifacts lead to misassignment of lysine ubiquitination sites by mass spectrometry.[8] Thus,there is aclear need for new electrophilic probes complementary to IAA that would enable labeling of so far "inaccessible" subsets of proteomic cysteines,which would then allow more comprehensive target profiling.Herein, we present discovery and an in-depth evaluation of a" clickable" alkynyl benziodoxolone as ahighly efficient and chemoselective cysteinereactive probe with an iodoacetamide-complementaryp r...

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A Direct Method for Site‐Specific Protein Acetylation

Cited by 128 publications

References 40 publications

Cytosolic Uptake of Large Monofunctionalized Dextrans

Cytosolic Uptake of Large Monofunctionalized Dextrans

Field Guide to Challenges and Opportunities in Antibody–Drug Conjugates for Chemists

Proteome‐Wide Profiling of Targets of Cysteine reactive Small Molecules by Using Ethynyl Benziodoxolone Reagents

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