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Hong Jiang et al.
Current protocols in chemical biology, 4(1), 19-34 (2012-11-29)
Poly(ADP-ribose) polymerases (PARPs) play important roles in various biological processes, including DNA repair, transcriptional regulation, mitosis, and RNA processing. PARP inhibitors are in clinical trials for treating human cancers. Understanding the biological function of PARPs will be important to fully
Emi Takamitsu et al.
Analytical biochemistry, 464, 83-93 (2014-07-22)
To establish a non-radioactive, cell-free detection system for protein N-myristoylation, metabolic labeling in a cell-free protein synthesis system using bioorthogonal myristic acid analogues was performed. After Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) with a biotin tag, the tagged proteins were separated by
Sandeep Ravindran et al.
PloS one, 4(12), e8143-e8143 (2009-12-04)
Toxoplasma gondii is a eukaryotic parasite of the phylum Apicomplexa that is able to infect a wide variety of host cells. During its active invasion process it secretes proteins from discrete secretory organelles: the micronemes, rhoptries and dense granules. Although
Caiping Tian et al.
Cell chemical biology, 24(11), 1416-1427 (2017-10-11)
Electrophilic groups, such as Michael acceptors, expoxides, are common motifs in natural products (NPs). Electrophilic NPs can act through covalent modification of cysteinyl thiols on functional proteins, and exhibit potent cytotoxicity and anti-inflammatory/cancer activities. Here we describe a new chemoproteomic
Yuanhui Ma et al.
Nature protocols, 13(8), 1744-1762 (2018-07-25)
Measuring proteome response to perturbations is critical for understanding the underlying mechanisms involved. Traditional quantitative proteomic methods are limited by the large numbers of proteins in the proteome and the mass spectrometer's dynamic range. A previous method uses the biorthogonal
Clive Yik-Sham Chung et al.
Nature chemical biology, 15(8), 776-785 (2019-07-10)
Autophagy is a lysosomal degradation pathway that eliminates aggregated proteins and damaged organelles to maintain cellular homeostasis. A major route for activating autophagy involves inhibition of the mTORC1 kinase, but current mTORC1-targeting compounds do not allow complete and selective mTORC1
Shaimaa M Aboukhatwa et al.
ChemMedChem, 14(11), 1096-1107 (2019-03-29)
Histone deacetylase (HDAC) activity is modulated in vivo by post-translational modifications and formation of multiprotein complexes. Novel chemical tools to study how these factors affect engagement of HDAC isoforms by HDAC inhibitors (HDACi) in cells and tissues are needed. In this
Mathias J Gerl et al.
PloS one, 11(4), e0153009-e0153009 (2016-04-23)
Cell membranes contain hundreds to thousands of individual lipid species that are of structural importance but also specifically interact with proteins. Due to their highly controlled synthesis and role in signaling events sphingolipids are an intensely studied class of lipids.
Rory K Morgan et al.
ACS chemical biology, 10(8), 1778-1784 (2015-05-16)
ADP-ribosylation is essential for cell function, yet there is a dearth of methods for detecting this post-translational modification in cells. Here, we describe a clickable aminooxy alkyne (AO-alkyne) probe that can detect cellular ADP-ribosylation on acidic amino acids following Cu-catalyzed
Susie J Zoltewicz et al.
ASN neuro, 4(6), 409-421 (2012-11-07)
PMP22 (peripheral myelin protein 22), also known as GAS 3 (growth-arrest-specific protein 3), is a disease-linked tetraspan glycoprotein of peripheral nerve myelin and constituent of intercellular junctions in epithelia. To date, our knowledge of the post-translational modification of PMP22 is
Ryuichiro Nakai et al.
Bioorganic & medicinal chemistry, 17(3), 1101-1108 (2008-03-28)
High-throughput screening (HTS) has become an integral part of academic and industrial efforts aimed at developing new chemical probes and drugs. These screens typically generate several 'hits', or lead active compounds, that must be prioritized for follow-up medicinal chemistry studies.
Bruce G Mockett et al.
The Journal of neuroscience : the official journal of the Society for Neuroscience, 39(17), 3188-3203 (2019-02-26)
Secreted amyloid precursor protein-alpha (sAPPα) has growth factor-like properties and can modulate long-term potentiation (LTP) and memory. Here, we demonstrate that exposure to sAPPα converts short-lasting LTP into protein-synthesis-dependent late LTP in hippocampal slices from male rats. sAPPβ had no
Michal Segal et al.
Biology open, 1(3), 220-231 (2012-12-06)
Bidirectional transport is a key issue in cellular biology. It requires coordination between microtubule-associated molecular motors that work in opposing directions. The major retrograde and anterograde motors involved in bidirectional transport are cytoplasmic dynein and conventional kinesin, respectively. It is
Teresa Galán et al.
Biosensors & bioelectronics, 74, 751-756 (2015-07-27)
Here we describe a label-free electrochemical DNA sensor based on poly(3,4-ethylenedioxythiophene)-modified (PEDOT-modified) electrodes. An acetylene-terminated DNA probe, complementary to a specific "Hepatitis C" virus sequence, was immobilized onto azido-derivatized conducting PEDOT electrodes using "click" chemistry. DNA hybridization was then detected
Eliot Morrison et al.
Communications biology, 3(1), 368-368 (2020-07-12)
Palmitoylation is the reversible addition of palmitate to cysteine via a thioester linkage. The reversible nature of this modification makes it a prime candidate as a mechanism for regulating signal transduction in T-cell receptor signaling. Following stimulation of the T-cell
Jung-Hyun Choi et al.
Molecular and cellular biology, 39(4) (2018-11-28)
Fragile X syndrome (FXS) caused by loss of fragile X mental retardation protein (FMRP), is the most common cause of inherited intellectual disability. Numerous studies show that FMRP is an RNA binding protein that regulates translation of its binding targets
Daniel W Bak et al.
Cell chemical biology, 25(9), 1157-1167 (2018-07-10)
Mammalian selenocysteine (Sec)-containing proteins, selenoproteins, are important to (patho)physiological processes, including redox homeostasis. Sec residues have been recalcitrant to mass spectrometry-based chemoproteomic methods that enrich for reactive cysteine (Cys) residues with electrophilic chemical probes, despite confirmed reactivity of Sec with
Lisa Roxanne Knoke et al.
Frontiers in microbiology, 11, 1228-1228 (2020-06-26)
Outer membrane vesicles (OMVs), released from Gram-negative bacteria, have been attributed to intra- and interspecies communication and pathogenicity in diverse bacteria. OMVs carry various components including genetic material, toxins, signaling molecules, or proteins. Although the molecular mechanism(s) of cargo delivery
Mingming Zhang et al.
Nature, 586(7829), 434-439 (2020-10-09)
Cysteine palmitoylation (S-palmitoylation) is a reversible post-translational modification that is installed by the DHHC family of palmitoyltransferases and is reversed by several acyl protein thioesterases1,2. Although thousands of human proteins are known to undergo S-palmitoylation, how this modification is regulated
Discovery of a Functional Covalent Ligand Targeting an Intrinsically Disordered Cysteine within MYC.
Lydia Boike et al.
Cell chemical biology, 28(1), 4-13 (2020-09-24)
MYC is a major oncogenic transcriptional driver of most human cancers that has remained intractable to direct targeting because much of MYC is intrinsically disordered. Here, we have performed a cysteine-reactive covalent ligand screen to identify compounds that could disrupt
Frank Iversen et al.
Theranostics, 3(3), 201-209 (2013-03-09)
Some of the main concerns with in vivo application of naked small interfering RNA are rapid degradation and urinary excretion resulting in a short plasma half-life. In this study we investigated how conjugation of polyethylene glycol (PEG) with variable chain
Chayasith Uttamapinant et al.
Nature protocols, 8(8), 1620-1634 (2013-07-28)
This protocol describes an efficient method to site-specifically label cell-surface or purified proteins with chemical probes in two steps: probe incorporation mediated by enzymes (PRIME) followed by chelation-assisted copper-catalyzed azide-alkyne cycloaddition (CuAAC). In the PRIME step, Escherichia coli lipoic acid
Thomas W Hanigan et al.
Cell chemical biology, 24(11), 1356-1367 (2017-09-26)
Histone deacetylase (HDAC) catalytic activity is regulated by formation of co-regulator complexes and post-translational modification. Whether these mechanisms are transformed in cancer and how this affects the binding and selectivity of HDAC inhibitors (HDACis) is unclear. In this study, we
Jung-Hyun Choi et al.
The EMBO journal, 37(5) (2018-02-15)
In neuronal development, dynamic rearrangement of actin promotes axonal growth cone extension, and spatiotemporal translation of local mRNAs in response to guidance cues directs axonal growth cone steering, where cofilin plays a critical role. While regulation of cofilin activity is
Saskia Heybrock et al.
Nature communications, 10(1), 3521-3521 (2019-08-08)
The intracellular transport of cholesterol is subject to tight regulation. The structure of the lysosomal integral membrane protein type 2 (LIMP-2, also known as SCARB2) reveals a large cavity that traverses the molecule and resembles the cavity in SR-B1 that
Yi-Jia Li et al.
Cell chemical biology, 26(2), 278-288 (2018-12-26)
Ubiquitin-like (Ubl) post-translational modifications are potential targets for therapeutics. However, the only known mechanism for inhibiting a Ubl-activating enzyme is through targeting its ATP-binding site. Here we identify an allosteric inhibitory site in the small ubiquitin-like modifier (SUMO)-activating enzyme (E1). This
Tilman Läppchen et al.
European journal of medicinal chemistry, 89, 279-295 (2014-12-03)
Calixarene 0118 is a potent anti-angiogenic agent that effectively inhibited tumor growth in preclinical studies, and is currently being evaluated in a phase I clinical trial. We have designed two close mimetics of calixarene 0118 containing a terminal alkynyl-functional group
Mark F van Delft et al.
Nature chemical biology, 15(11), 1057-1066 (2019-10-09)
Activating the intrinsic apoptosis pathway with small molecules is now a clinically validated approach to cancer therapy. In contrast, blocking apoptosis to prevent the death of healthy cells in disease settings has not been achieved. Caspases have been favored, but
Migle Tomkuviene et al.
Nucleic acids research, 40(14), 6765-6773 (2012-05-09)
Biophysical and mechanistic investigation of RNA function requires site-specific incorporation of spectroscopic and chemical probes, which is difficult to achieve using current technologies. We have in vitro reconstituted a functional box C/D small ribonucleoprotein RNA 2'-O-methyltransferase (C/D RNP) from the
Keli Chai et al.
Viruses, 13(1) (2020-12-31)
Membrane proteins of enveloped viruses have been reported to undergo palmitoylation, a post-translational modification often having a critical role in the function of these viral proteins and hence viral replication. In this study, we report that the foamy virus (FV)
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