Biosensors Show the Pharmacokinetics of S-Ketamine in the Endoplasmic Reticulum
The target for the “rapid” (<24 h) antidepressant effects of S-ketamine is unknown, vitiating programs to rationally develop more effective rapid antidepressants. To describe a drug’s target, one must first understand the compartments entered by the drug, at all levels—the organ, the cell, an...
| Main Authors: | , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2019-11-01
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| Series: | Frontiers in Cellular Neuroscience |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/article/10.3389/fncel.2019.00499/full |
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| author | Kallol Bera Aron Kamajaya Amol V. Shivange Anand K. Muthusamy Anand K. Muthusamy Aaron L. Nichols Aaron L. Nichols Philip M. Borden Stephen Grant Janice Jeon Elaine Lin Ishak Bishara Theodore M. Chin Bruce N. Cohen Charlene H. Kim Elizabeth K. Unger Lin Tian Jonathan S. Marvin Loren L. Looger Henry A. Lester |
| author_facet | Kallol Bera Aron Kamajaya Amol V. Shivange Anand K. Muthusamy Anand K. Muthusamy Aaron L. Nichols Aaron L. Nichols Philip M. Borden Stephen Grant Janice Jeon Elaine Lin Ishak Bishara Theodore M. Chin Bruce N. Cohen Charlene H. Kim Elizabeth K. Unger Lin Tian Jonathan S. Marvin Loren L. Looger Henry A. Lester |
| author_sort | Kallol Bera |
| collection | DOAJ |
| description | The target for the “rapid” (<24 h) antidepressant effects of S-ketamine is unknown, vitiating programs to rationally develop more effective rapid antidepressants. To describe a drug’s target, one must first understand the compartments entered by the drug, at all levels—the organ, the cell, and the organelle. We have, therefore, developed molecular tools to measure the subcellular, organellar pharmacokinetics of S-ketamine. The tools are genetically encoded intensity-based S-ketamine-sensing fluorescent reporters, iSKetSnFR1 and iSKetSnFR2. In solution, these biosensors respond to S-ketamine with a sensitivity, S-slope = delta(F/F0)/(delta[S-ketamine]) of 0.23 and 1.9/μM, respectively. The iSKetSnFR2 construct allows measurements at <0.3 μM S-ketamine. The iSKetSnFR1 and iSKetSnFR2 biosensors display >100-fold selectivity over other ligands tested, including R-ketamine. We targeted each of the sensors to either the plasma membrane (PM) or the endoplasmic reticulum (ER). Measurements on these biosensors expressed in Neuro2a cells and in human dopaminergic neurons differentiated from induced pluripotent stem cells (iPSCs) show that S-ketamine enters the ER within a few seconds after appearing in the external solution near the PM, then leaves as rapidly after S-ketamine is removed from the extracellular solution. In cells, S-slopes for the ER and PM-targeted sensors differ by <2-fold, indicating that the ER [S-ketamine] is less than 2-fold different from the extracellular [S-ketamine]. Organelles represent potential compartments for the engagement of S-ketamine with its antidepressant target, and potential S-ketamine targets include organellar ion channels, receptors, and transporters. |
| first_indexed | 2024-12-21T19:05:25Z |
| format | Article |
| id | doaj.art-4397d649b8754ebaa97c121f59eaa73b |
| institution | Directory Open Access Journal |
| issn | 1662-5102 |
| language | English |
| last_indexed | 2024-12-21T19:05:25Z |
| publishDate | 2019-11-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular Neuroscience |
| spelling | doaj.art-4397d649b8754ebaa97c121f59eaa73b2022-12-21T18:53:21ZengFrontiers Media S.A.Frontiers in Cellular Neuroscience1662-51022019-11-011310.3389/fncel.2019.00499492298Biosensors Show the Pharmacokinetics of S-Ketamine in the Endoplasmic ReticulumKallol Bera0Aron Kamajaya1Amol V. Shivange2Anand K. Muthusamy3Anand K. Muthusamy4Aaron L. Nichols5Aaron L. Nichols6Philip M. Borden7Stephen Grant8Janice Jeon9Elaine Lin10Ishak Bishara11Theodore M. Chin12Bruce N. Cohen13Charlene H. Kim14Elizabeth K. Unger15Lin Tian16Jonathan S. Marvin17Loren L. Looger18Henry A. Lester19Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, United StatesDivision of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, United StatesDivision of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, United StatesDivision of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, United StatesDivision of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA, United StatesDivision of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, United StatesDivision of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA, United StatesJanelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, United StatesDivision of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA, United StatesDivision of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, United StatesDivision of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, United StatesDivision of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, United StatesDivision of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, United StatesDivision of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, United StatesDivision of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, United StatesDepartment of Biochemistry and Molecular Medicine, University of California, Davis, Davis, CA, United StatesDepartment of Biochemistry and Molecular Medicine, University of California, Davis, Davis, CA, United StatesJanelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, United StatesJanelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, United StatesDivision of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, United StatesThe target for the “rapid” (<24 h) antidepressant effects of S-ketamine is unknown, vitiating programs to rationally develop more effective rapid antidepressants. To describe a drug’s target, one must first understand the compartments entered by the drug, at all levels—the organ, the cell, and the organelle. We have, therefore, developed molecular tools to measure the subcellular, organellar pharmacokinetics of S-ketamine. The tools are genetically encoded intensity-based S-ketamine-sensing fluorescent reporters, iSKetSnFR1 and iSKetSnFR2. In solution, these biosensors respond to S-ketamine with a sensitivity, S-slope = delta(F/F0)/(delta[S-ketamine]) of 0.23 and 1.9/μM, respectively. The iSKetSnFR2 construct allows measurements at <0.3 μM S-ketamine. The iSKetSnFR1 and iSKetSnFR2 biosensors display >100-fold selectivity over other ligands tested, including R-ketamine. We targeted each of the sensors to either the plasma membrane (PM) or the endoplasmic reticulum (ER). Measurements on these biosensors expressed in Neuro2a cells and in human dopaminergic neurons differentiated from induced pluripotent stem cells (iPSCs) show that S-ketamine enters the ER within a few seconds after appearing in the external solution near the PM, then leaves as rapidly after S-ketamine is removed from the extracellular solution. In cells, S-slopes for the ER and PM-targeted sensors differ by <2-fold, indicating that the ER [S-ketamine] is less than 2-fold different from the extracellular [S-ketamine]. Organelles represent potential compartments for the engagement of S-ketamine with its antidepressant target, and potential S-ketamine targets include organellar ion channels, receptors, and transporters.https://www.frontiersin.org/article/10.3389/fncel.2019.00499/fullantidepressantsorganellesgreen fluorescent proteinprotein engineering and designperiplasmic binding proteins (PBPs)inside-out pharmacology |
| spellingShingle | Kallol Bera Aron Kamajaya Amol V. Shivange Anand K. Muthusamy Anand K. Muthusamy Aaron L. Nichols Aaron L. Nichols Philip M. Borden Stephen Grant Janice Jeon Elaine Lin Ishak Bishara Theodore M. Chin Bruce N. Cohen Charlene H. Kim Elizabeth K. Unger Lin Tian Jonathan S. Marvin Loren L. Looger Henry A. Lester Biosensors Show the Pharmacokinetics of S-Ketamine in the Endoplasmic Reticulum Frontiers in Cellular Neuroscience antidepressants organelles green fluorescent protein protein engineering and design periplasmic binding proteins (PBPs) inside-out pharmacology |
| title | Biosensors Show the Pharmacokinetics of S-Ketamine in the Endoplasmic Reticulum |
| title_full | Biosensors Show the Pharmacokinetics of S-Ketamine in the Endoplasmic Reticulum |
| title_fullStr | Biosensors Show the Pharmacokinetics of S-Ketamine in the Endoplasmic Reticulum |
| title_full_unstemmed | Biosensors Show the Pharmacokinetics of S-Ketamine in the Endoplasmic Reticulum |
| title_short | Biosensors Show the Pharmacokinetics of S-Ketamine in the Endoplasmic Reticulum |
| title_sort | biosensors show the pharmacokinetics of s ketamine in the endoplasmic reticulum |
| topic | antidepressants organelles green fluorescent protein protein engineering and design periplasmic binding proteins (PBPs) inside-out pharmacology |
| url | https://www.frontiersin.org/article/10.3389/fncel.2019.00499/full |
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