Abstract:
:Phenotypic screening of compound libraries is a significant trend in drug discovery, yet success can be hindered by difficulties in identifying the underlying cellular targets. Current approaches rely on tethering bioactive compounds to a capture tag or surface to allow selective enrichment of interacting proteins for subsequent identification by mass spectrometry. Such methods are often constrained by ineffective capture of low affinity and low abundance targets. In addition, these methods are often not compatible with living cells and therefore cannot be used to verify the pharmacological activity of the tethered compounds. We have developed a novel chloroalkane capture tag that minimally affects compound potency in cultured cells, allowing binding interactions with the targets to occur under conditions relevant to the desired cellular phenotype. Subsequent isolation of the interacting targets is achieved through rapid lysis and capture onto immobilized HaloTag protein. Exchanging the chloroalkane tag for a fluorophore, the putative targets identified by mass spectrometry can be verified for direct binding to the compound through resonance energy transfer. Using the interaction between histone deacetylases (HDACs) and the inhibitor, Vorinostat (SAHA), as a model system, we were able to identify and verify all the known HDAC targets of SAHA as well as two previously undescribed targets, ADO and CPPED1. The discovery of ADO as a target may provide mechanistic insight into a reported connection between SAHA and Huntington's disease.
journal_name
ACS Chem Bioljournal_title
ACS chemical biologyauthors
Ohana RF,Kirkland TA,Woodroofe CC,Levin S,Uyeda HT,Otto P,Hurst R,Robers MB,Zimmerman K,Encell LP,Wood KVdoi
10.1021/acschembio.5b00351subject
Has Abstractpub_date
2015-10-16 00:00:00pages
2316-24issue
10eissn
1554-8929issn
1554-8937journal_volume
10pub_type
杂志文章abstract::Human glucosylcerebrosidase 2 (GBA2) of the CAZy family GH116 is responsible for the breakdown of glycosphingolipids on the cytoplasmic face of the endoplasmic reticulum and Golgi apparatus. Genetic defects in GBA2 result in spastic paraplegia and cerebellar ataxia, while cross-talk between GBA2 and GBA1 glucosylceram...
journal_title:ACS chemical biology
pub_type: 杂志文章
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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
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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
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journal_title:ACS chemical biology
pub_type: 杂志文章
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更新日期:2017-11-17 00:00:00
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journal_title:ACS chemical biology
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journal_title:ACS chemical biology
pub_type: 信件
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journal_title:ACS chemical biology
pub_type: 杂志文章
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journal_title:ACS chemical biology
pub_type: 杂志文章
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journal_title:ACS chemical biology
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pub_type: 传,历史文章,杂志文章
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journal_title:ACS chemical biology
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journal_title:ACS chemical biology
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journal_title:ACS chemical biology
pub_type: 杂志文章
doi:10.1021/cb6003788
更新日期:2007-02-20 00:00:00
abstract::In a process called quorum sensing, bacteria produce and secrete certain signaling compounds (called autoinducers) that bind to receptors on other bacteria and activate transcription of certain genes. A clever genetic selection yields a new quorum-sensing transcriptional regulator that marches to the beat of a differe...
journal_title:ACS chemical biology
pub_type: 杂志文章,评审
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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::Fusicoccin A is a diterpene glucoside phytotoxin generated by the fungal pathogen Phomopsis amygdali that causes the plant disease constriction canker, first discovered in New Jersey peach orchards in the 1930s. Fusicoccin A is also an emerging new lead in cancer chemotherapy. The hydrocarbon precursor of fusicoccin A...
journal_title:ACS chemical biology
pub_type: 杂志文章
doi:10.1021/acschembio.5b00960
更新日期:2016-04-15 00:00:00
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journal_title:ACS chemical biology
pub_type: 杂志文章
doi:10.1021/acschembio.8b00952
更新日期:2019-02-15 00:00:00
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journal_title:ACS chemical biology
pub_type: 杂志文章
doi:10.1021/acschembio.7b00060
更新日期:2017-05-19 00:00:00
abstract::Nearly one-third of the encoded proteome is comprised of secretory proteins that enable communication between cells and organ systems, playing a ubiquitous role in human health and disease. High-throughput detection of secreted proteins would enhance efforts to identify therapies for secretion-related diseases. Using ...
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doi:10.1021/acschembio.9b00001
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更新日期:2015-09-18 00:00:00
abstract::Ubc13 is an E2 ubiquitin conjugating enzyme that functions in nuclear DNA damage signaling and cytoplasmic NF-κB signaling. Here, we present the structures of complexes of Ubc13 with two inhibitors, NSC697923 and BAY 11-7082, which inhibit DNA damage and NF-κB signaling in human cells. NSC697923 and BAY 11-7082 both i...
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更新日期:2021-01-15 00:00:00
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journal_title:ACS chemical biology
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abstract::Caenorhabditis elegans lives in compost and decaying fruit, eats bacteria and is exposed to pathogenic microbes. We show that C. elegans is able to modify diverse microbial small-molecule toxins via both O- and N-glucosylation as well as unusual 3'-O-phosphorylation of the resulting glucosides. The resulting glucosyla...
journal_title:ACS chemical biology
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abstract::Seasonal and pandemic influenza viruses continue to be a leading global health concern. Emerging resistance to the current drugs and the variable efficacy of vaccines underscore the need for developing new flu drugs that will be broadly effective against wild-type and drug-resistant influenza strains. Here, we report ...
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journal_title:ACS chemical biology
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