Phenelzine-based probes reveal Secernin-3 is involved in thermal nociception

Bustin, Katelyn A.; Shishikura, Kyosuke; Chen, Irene; Lin, Zongtao; McKnight, Nate R; Chang, Yuxuan; Wang, Xie; Li, Jing Jing; Arellano, Eric; Pei, Liming; Morton, Paul D.; Gregus, Ann M.; Buczynski, Matthew W.; Matthews, Megan L.

doi:10.1016/j.mcn.2023.103842

Cited by 3 publications

(2 citation statements)

References 74 publications

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“…The speed of chemical proteomics is highly dependent on the manual process of sample preparation ,,− and megadata analysis. ,− While data processing has been greatly accelerated by computational technologies, manual sample preparation remains the major bottleneck of high-throughput chemical proteomics in drug target discovery. Currently, most sample preparation protocols require an exhaustive and systematic series of manual steps that must be carried out by skilled scientists to extract the proteome fraction of interest (e.g., a probe-labeled proteome for chemical proteomics). − ,− For many protocols, sample preparations require days of labor and inflexible hours at the bench due to the inability to break down the steps and the need to maintain consistency. It can take months of repetitive bench work to complete one data set from proteomics samples with a maximum throughput of ∼10 samples/week/scientist.…”

Section: Introductionmentioning

confidence: 99%

“…This approach has enabled new opportunities for the discovery of protein drug targets, protein modifications, and small-molecule drugs that selectively modulate their functions (Figure ). − There are large numbers of chemical libraries available with untapped potential for new discoveries, each with unique reactivities across diverse biological samples (cell lysates, cell cultures or populations, tissues, organs, and whole organisms). − Chemical proteomics often employs a variety of preparations for drug target and drug mechanism discovery. ,,− Many protocols require multiple operational steps and often share common processing steps. ,− A short list of common processes includes cell lysis, chemical reactions, purification or fractionation of a proteome of interest (POI), protein denaturation, reduction and alkylation of protein thiols, enzymatic proteolytic digestion of proteins followed by isotopic labeling with chemical tags, and peptide desalting. The speed of chemical proteomics is highly dependent on the manual process of sample preparation ,,− and megadata analysis. ,− While data processing has been greatly accelerated by computational technologies, manual sample preparation remains the major bottleneck of high-throughput chemical proteomics in drug target discovery.…”

Section: Introductionmentioning

confidence: 99%

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Automation to Enable High-Throughput Chemical Proteomics

Lin,

Gongora,

Liu

et al. 2023

J. Proteome Res.

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Chemical proteomics utilizes small-molecule probes to covalently engage with their interacting proteins. Since chemical probes are tagged to the active or binding sites of functional proteins, chemical proteomics can be used to profile protein targets, reveal precise binding sites/mechanisms, and screen inhibitors competing with probes in a biological context. These capabilities of chemical proteomics have great potential to enable discoveries of both drug targets and lead compounds. However, chemical proteomics is limited by the time-consuming bottleneck of sample preparations, which are processed manually. With the advancement of robotics and artificial intelligence, it is now possible to automate workflows to make chemical proteomics sample preparation a high-throughput process. An automated robotic system represents a major technological opportunity to speed up advances in proteomics, open new frontiers in drug target discovery, and broaden the future of chemical biology.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Automation to Enable High-Throughput Chemical Proteomics

Lin,

Gongora,

Liu

et al. 2023

J. Proteome Res.

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Genetics and epigenetics approaches as a path to the future of addiction science

West,

Day

2023

Molecular and Cellular Neuroscience

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12/15-Lipoxygenases mediate neuropathic-like pain hypersensitivity in female mice

Brown,

Chen,

Miliano

et al. 2024

Preprint

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It is estimated that chronic neuropathic pain conditions exhibit up to 10% prevalence in the general population, with increased incidence in females. However, nonsteroidal inflammatory drugs (NSAIDs) are ineffective, and currently indicated prescription treatments such as opioids, anticonvulsants, and antidepressants provide only limited therapeutic benefit. In the current work, we extended previous studies in male rats utilizing a paradigm of central Toll-like receptor 4 (TLR4)-dependent, NSAID-unresponsive neuropathic-like pain hypersensitivity to male and female C57BL/6N mice, uncovering an unexpected hyperalgesic phenotype in female mice following intrathecal (IT) LPS. In contrast to previous reports in female C57BL/6J mice, female C57BL/6N mice displayed tactile and cold allodynia, grip force deficits, and locomotor hyperactivity in response to IT LPS. Congruent with our previous observations in male rats, systemic inhibition of 12/15-Lipoxygenases (12/15-LOX) in female B6N mice with selective inhibitors - ML355 (targeting 12-LOX-p) and ML351 (targeting 15-LOX-1) - completely reversed allodynia and grip force deficits. We demonstrate here that 12/15-LOX enzymes also are expressed in mouse spinal cord and that 12/15-LOX metabolites produce tactile allodynia when administered spinally (IT) or peripherally (intraplantar in the paw, IPLT) in a hyperalgesic priming model, similar to others observations with the cyclooxygenase (COX) metabolite Prostaglandin E2 (PGE2). Surprisingly, we did not detect hyperalgesic priming following IT administration of LPS, indicating that this phenomenon likely requires peripheral activation of nociceptors. Collectively, these data suggest that 12/15-LOX enzymes contribute to neuropathic-like pain hypersensitivity in rodents, with potential translatability as druggable targets across sexes and species using multiple reflexive and non-reflexive outcome measures.

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Phenelzine-based probes reveal Secernin-3 is involved in thermal nociception

Cited by 3 publications

References 74 publications

Automation to Enable High-Throughput Chemical Proteomics

Automation to Enable High-Throughput Chemical Proteomics

Genetics and epigenetics approaches as a path to the future of addiction science

12/15-Lipoxygenases mediate neuropathic-like pain hypersensitivity in female mice

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