Get 20M+ Full-Text Papers For Less Than $1.50/day. Start a 14-Day Trial for You or Your Team.

Learn More →

High-throughput screening reveals enzyme and GPCR targets as putative binding sites for D-deprenyl

High-throughput screening reveals enzyme and GPCR targets as putative binding sites for D-deprenyl 52 Abstracts / Scandinavian Journal of Pain 8 (2015) 47–54 High-throughput screening reveals enzyme and New players in the mechanism of spinal cord GPCR targets as putative binding sites for stimulation for neuropathic pain d-deprenyl a,∗ b c a A. Lind , P. Emami , M. Sjödin , L. Katila ,M. a,∗ a b c a b A. Lesniak , A. Jonsson , M. Aarnio ,T. Wetterhall , T. Gordh , K. Kultima c a b Norberg , F. Nyberg , T. Gordh Department of Surgical Sciences, Anaesthesiology Uppsala University, Department of Pharmaceutical and Intensive Care, Uppsala University, SE-75185 Biosciences, Uppsala, Sweden Uppsala, Sweden b b Uppsala University Hospital, Department of Department of Medical Sciences, Cancer Surgical Sciences, Anaesthesiology and Intensive Pharmacology and Computational Medicine, Care, Uppsala, Sweden Uppsala University, SE-75185 Uppsala, Sweden c c Uppsala University, Department of Chemistry, Department of Chemistry-BMC, Analytical Uppsala, Sweden Chemistry, Uppsala University, GE Healthcare, E-mail address: anna.lesniak@farmbio.uu.se (A. SE-75124, Uppsala, Sweden Lesniak). E-mail address: anne-li.lind@surgsci.uu.se (A. Lind). Aims: In PET studies of patients suffering from chronic pain fol- lowing whip lash trauma, d-deprenyl was shown to bind to painful Aims: The aim of this study was to uncover possible proteins sites in the neck [1]. High uptake points towards an existence of involved in the mechanism for neuropathic spinal cord stimulation an inflammation-specific binding site. Thus, the aim of this study (SCS) pain relief in humans. was to identify the binding site for d-deprenyl employing radioli- Methods: Using two different proteomic protocols we com- gand receptor binding and high-throughput analysis of its activity pared the protein concentration in cerebrospinal fluid (CSF) from towards 165 G-protein coupled receptors and 84 enzyme targets 14 SCS responsive neuropathic pain patients using a shotgun pro- commonly used in drug discovery and development. teomic approach. The comparison was made between samples from Methods: d-Deprenyl activity towards GPCR targets was the same individual taken when the stimulator had been off for 48 h, assessed by DiscoverX in CHO-K1 EDG1 -arrestin EFC cell line and after the stimulator had been used as normal for three weeks. TM utilizing the PathHunter technique. Enzyme inhibition by D- Mass spectrometry raw files from both experimental setups were TM deprenyl was identified in the EnzymeProfiling screening panel analysed using MS Quant and DeCyder softwares and proteins were provided by Eurofins Cerep Panlabs. [H ]d-deprenyl binding stud- identified using MASCOT search against Swissprot database. After ies with specific GPCR agonists and enzyme inhibitors at newly median normalization moderate paired t-test statistics were used identified targets were also performed. in R limma package to find up-and down regulated proteins. Results: Our investigation revealed that a 10M concentra- Results: In total, 255 and 216 proteins could be identified by the tion of d-deprenyl inhibited MAO-B and MAO-A activity by 99% dimethyl and label free methods respectively and relatively quan- and 55%, respectively. In addition, a 70% inhibition of angiotensin tified. Of these several were significantly up- and downregulated. converting enzyme (ACE) activity in rabbit lung preparations was Conclusions: These novel proteins add to the SCS mechanism found. Furthermore, binding studies in rat mitochondrial mem- in patients with neuropathic pain. These results strengthen the brane homogenates confirmed a submicromolar [H ]d-deprenyl evidence for SCS as altering extracellular matrix components and competition with a selective MAO-B inhibitor seligiline, but not affecting central nervous system plasticity. with the selective MAO-A inhibitor pirlindole mesylate. No evident hits among GPCR targets were identified. However, attention was http://dx.doi.org/10.1016/j.sjpain.2015.04.017 drawn towards the histamine HRH1 and HRH3 receptors to which d-deprenyl showed a 20% and 42% antagonistic activity. Hyperalgesia after experimental and Conclusions: MAO-B might be a candidate target ford-deprenyl, work-related sleep restriction as many other studies documented higher d-deprenyl uptake in a,b,c a,∗ K.B. Nilsen , D. Matre activated astrocytes, non-secreting pituitary adenomas and brown adipose tissue, where MAO-B is overexpressed. Moreover, ACE National Institute of Occupational Health, inhibition was shown to hamper down-regulation of transcription Department of Work Psychology and Physiology, factors preventing ROS-mediated cartilage damage. Oslo, Norway Norwegian University of Science and Technology, Reference Department of Neuroscience, Trondheim, Norway Oslo University Hospital – Ullevål, Department of [1] Linnman C, Appel L, Fredrikson M, Gordh T, Soderlund A, Langstrom B, Engler H. Elevated [ C]-d-deprenyl uptake in chronic Whiplash Associated Disorder Neurology, Section for Clinical Neurophysiology, suggests persistent musculoskeletal inflammation. PLoS ONE 2011;6:e19182. Norway E-mail address: dagfinn.matre@stami.no (D. http://dx.doi.org/10.1016/j.sjpain.2015.04.016 Matre). Aims: Sleep restriction (SR) increases pain sensitivity. The aim of this study was to compare the effects of night work on pain sensitivity with experimental SR. Methods: In study I 22 healthy volunteers (14 females) and in study II 24 nurses (17 females) received pain stimuli in the labo- ratory twice; after 2 nights with habitual sleep (study I and II) and after 2 nights of experimental 50% SR (study I) or after 2 nights of work (study II). Order of sleep conditions was randomized. Heat pain at intensity 6/10 (pain-6), the pressure pain threshold (PPT), and subjective sleepiness with Karolinska sleepiness scale (KSS) http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Scandinavian Journal of Pain de Gruyter

High-throughput screening reveals enzyme and GPCR targets as putative binding sites for D-deprenyl

Download PDF
1 page

Loading next page...
 
/lp/degruyter/high-throughput-screening-reveals-enzyme-and-gpcr-targets-as-putative-AHsXoAipTy

References

References for this paper are not available at this time. We will be adding them shortly, thank you for your patience.

Publisher
de Gruyter
Copyright
© 2015 Scandinavian Association for the Study of Pain
ISSN
1877-8860
eISSN
1877-8879
DOI
10.1016/j.sjpain.2015.04.016
Publisher site
See Article on Publisher Site

Abstract

52 Abstracts / Scandinavian Journal of Pain 8 (2015) 47–54 High-throughput screening reveals enzyme and New players in the mechanism of spinal cord GPCR targets as putative binding sites for stimulation for neuropathic pain d-deprenyl a,∗ b c a A. Lind , P. Emami , M. Sjödin , L. Katila ,M. a,∗ a b c a b A. Lesniak , A. Jonsson , M. Aarnio ,T. Wetterhall , T. Gordh , K. Kultima c a b Norberg , F. Nyberg , T. Gordh Department of Surgical Sciences, Anaesthesiology Uppsala University, Department of Pharmaceutical and Intensive Care, Uppsala University, SE-75185 Biosciences, Uppsala, Sweden Uppsala, Sweden b b Uppsala University Hospital, Department of Department of Medical Sciences, Cancer Surgical Sciences, Anaesthesiology and Intensive Pharmacology and Computational Medicine, Care, Uppsala, Sweden Uppsala University, SE-75185 Uppsala, Sweden c c Uppsala University, Department of Chemistry, Department of Chemistry-BMC, Analytical Uppsala, Sweden Chemistry, Uppsala University, GE Healthcare, E-mail address: anna.lesniak@farmbio.uu.se (A. SE-75124, Uppsala, Sweden Lesniak). E-mail address: anne-li.lind@surgsci.uu.se (A. Lind). Aims: In PET studies of patients suffering from chronic pain fol- lowing whip lash trauma, d-deprenyl was shown to bind to painful Aims: The aim of this study was to uncover possible proteins sites in the neck [1]. High uptake points towards an existence of involved in the mechanism for neuropathic spinal cord stimulation an inflammation-specific binding site. Thus, the aim of this study (SCS) pain relief in humans. was to identify the binding site for d-deprenyl employing radioli- Methods: Using two different proteomic protocols we com- gand receptor binding and high-throughput analysis of its activity pared the protein concentration in cerebrospinal fluid (CSF) from towards 165 G-protein coupled receptors and 84 enzyme targets 14 SCS responsive neuropathic pain patients using a shotgun pro- commonly used in drug discovery and development. teomic approach. The comparison was made between samples from Methods: d-Deprenyl activity towards GPCR targets was the same individual taken when the stimulator had been off for 48 h, assessed by DiscoverX in CHO-K1 EDG1 -arrestin EFC cell line and after the stimulator had been used as normal for three weeks. TM utilizing the PathHunter technique. Enzyme inhibition by D- Mass spectrometry raw files from both experimental setups were TM deprenyl was identified in the EnzymeProfiling screening panel analysed using MS Quant and DeCyder softwares and proteins were provided by Eurofins Cerep Panlabs. [H ]d-deprenyl binding stud- identified using MASCOT search against Swissprot database. After ies with specific GPCR agonists and enzyme inhibitors at newly median normalization moderate paired t-test statistics were used identified targets were also performed. in R limma package to find up-and down regulated proteins. Results: Our investigation revealed that a 10M concentra- Results: In total, 255 and 216 proteins could be identified by the tion of d-deprenyl inhibited MAO-B and MAO-A activity by 99% dimethyl and label free methods respectively and relatively quan- and 55%, respectively. In addition, a 70% inhibition of angiotensin tified. Of these several were significantly up- and downregulated. converting enzyme (ACE) activity in rabbit lung preparations was Conclusions: These novel proteins add to the SCS mechanism found. Furthermore, binding studies in rat mitochondrial mem- in patients with neuropathic pain. These results strengthen the brane homogenates confirmed a submicromolar [H ]d-deprenyl evidence for SCS as altering extracellular matrix components and competition with a selective MAO-B inhibitor seligiline, but not affecting central nervous system plasticity. with the selective MAO-A inhibitor pirlindole mesylate. No evident hits among GPCR targets were identified. However, attention was http://dx.doi.org/10.1016/j.sjpain.2015.04.017 drawn towards the histamine HRH1 and HRH3 receptors to which d-deprenyl showed a 20% and 42% antagonistic activity. Hyperalgesia after experimental and Conclusions: MAO-B might be a candidate target ford-deprenyl, work-related sleep restriction as many other studies documented higher d-deprenyl uptake in a,b,c a,∗ K.B. Nilsen , D. Matre activated astrocytes, non-secreting pituitary adenomas and brown adipose tissue, where MAO-B is overexpressed. Moreover, ACE National Institute of Occupational Health, inhibition was shown to hamper down-regulation of transcription Department of Work Psychology and Physiology, factors preventing ROS-mediated cartilage damage. Oslo, Norway Norwegian University of Science and Technology, Reference Department of Neuroscience, Trondheim, Norway Oslo University Hospital – Ullevål, Department of [1] Linnman C, Appel L, Fredrikson M, Gordh T, Soderlund A, Langstrom B, Engler H. Elevated [ C]-d-deprenyl uptake in chronic Whiplash Associated Disorder Neurology, Section for Clinical Neurophysiology, suggests persistent musculoskeletal inflammation. PLoS ONE 2011;6:e19182. Norway E-mail address: dagfinn.matre@stami.no (D. http://dx.doi.org/10.1016/j.sjpain.2015.04.016 Matre). Aims: Sleep restriction (SR) increases pain sensitivity. The aim of this study was to compare the effects of night work on pain sensitivity with experimental SR. Methods: In study I 22 healthy volunteers (14 females) and in study II 24 nurses (17 females) received pain stimuli in the labo- ratory twice; after 2 nights with habitual sleep (study I and II) and after 2 nights of experimental 50% SR (study I) or after 2 nights of work (study II). Order of sleep conditions was randomized. Heat pain at intensity 6/10 (pain-6), the pressure pain threshold (PPT), and subjective sleepiness with Karolinska sleepiness scale (KSS)

Journal

Scandinavian Journal of Pain – de Gruyter

Published: Jul 1, 2015

There are no references for this article.