[{"publication":"Journal of Biological Chemistry","type":"journal_article","date_updated":"2025-04-14T07:44:00Z","scopus_import":"1","ddc":["570"],"article_type":"original","article_processing_charge":"No","year":"2022","volume":298,"title":"Depletion of endogenously biotinylated carboxylases enhances the sensitivity of TurboID-mediated proximity labeling in Caenorhabditis elegans","oa":1,"day":"01","abstract":[{"text":"Proximity-dependent protein labeling provides a powerful in vivo strategy to characterize the interactomes of specific proteins. We previously optimized a proximity labeling protocol for Caenorhabditis elegans using the highly active biotin ligase TurboID. A significant constraint on the sensitivity of TurboID is the presence of abundant endogenously biotinylated proteins that take up bandwidth in the mass spectrometer, notably carboxylases that use biotin as a cofactor. In C. elegans, these comprise POD-2/acetyl-CoA carboxylase alpha, PCCA-1/propionyl-CoA carboxylase alpha, PYC-1/pyruvate carboxylase, and MCCC-1/methylcrotonyl-CoA carboxylase alpha. Here, we developed ways to remove these carboxylases prior to streptavidin purification and mass spectrometry by engineering their corresponding genes to add a C-terminal His10 tag. This allows us to deplete them from C. elegans lysates using immobilized metal affinity chromatography. To demonstrate the method's efficacy, we use it to expand the interactome map of the presynaptic active zone protein ELKS-1. We identify many known active zone proteins, including UNC-10/RIM, SYD-2/liprin-alpha, SAD-1/BRSK1, CLA-1/CLArinet, C16E9.2/Sentryn, as well as previously uncharacterized potentially synaptic proteins such as the ortholog of human angiomotin, F59C12.3 and the uncharacterized protein R148.3. Our approach provides a quick and inexpensive solution to a common contaminant problem in biotin-dependent proximity labeling. The approach may be applicable to other model organisms and will enable deeper and more complete analysis of interactors for proteins of interest.","lang":"eng"}],"corr_author":"1","acknowledged_ssus":[{"_id":"Bio"}],"language":[{"iso":"eng"}],"article_number":"102343","ec_funded":1,"file_date_updated":"2022-09-12T08:14:50Z","publication_status":"published","doi":"10.1016/j.jbc.2022.102343","date_published":"2022-09-01T00:00:00Z","project":[{"grant_number":"209504/A/17/Z","_id":"23870BE8-32DE-11EA-91FC-C7463DDC885E","name":"Molecular mechanisms of neural circuit function"},{"grant_number":"754411","call_identifier":"H2020","name":"ISTplus - Postdoctoral Fellowships","_id":"260C2330-B435-11E9-9278-68D0E5697425"}],"quality_controlled":"1","_id":"12082","publication_identifier":{"eissn":["1083-351X"],"issn":["0021-9258"]},"intvolume":"       298","issue":"9","publisher":"Elsevier","citation":{"apa":"Artan, M., Hartl, M., Chen, W., &#38; de Bono, M. (2022). Depletion of endogenously biotinylated carboxylases enhances the sensitivity of TurboID-mediated proximity labeling in Caenorhabditis elegans. <i>Journal of Biological Chemistry</i>. Elsevier. <a href=\"https://doi.org/10.1016/j.jbc.2022.102343\">https://doi.org/10.1016/j.jbc.2022.102343</a>","ama":"Artan M, Hartl M, Chen W, de Bono M. Depletion of endogenously biotinylated carboxylases enhances the sensitivity of TurboID-mediated proximity labeling in Caenorhabditis elegans. <i>Journal of Biological Chemistry</i>. 2022;298(9). doi:<a href=\"https://doi.org/10.1016/j.jbc.2022.102343\">10.1016/j.jbc.2022.102343</a>","ieee":"M. Artan, M. Hartl, W. Chen, and M. de Bono, “Depletion of endogenously biotinylated carboxylases enhances the sensitivity of TurboID-mediated proximity labeling in Caenorhabditis elegans,” <i>Journal of Biological Chemistry</i>, vol. 298, no. 9. Elsevier, 2022.","ista":"Artan M, Hartl M, Chen W, de Bono M. 2022. Depletion of endogenously biotinylated carboxylases enhances the sensitivity of TurboID-mediated proximity labeling in Caenorhabditis elegans. Journal of Biological Chemistry. 298(9), 102343.","mla":"Artan, Murat, et al. “Depletion of Endogenously Biotinylated Carboxylases Enhances the Sensitivity of TurboID-Mediated Proximity Labeling in Caenorhabditis Elegans.” <i>Journal of Biological Chemistry</i>, vol. 298, no. 9, 102343, Elsevier, 2022, doi:<a href=\"https://doi.org/10.1016/j.jbc.2022.102343\">10.1016/j.jbc.2022.102343</a>.","short":"M. Artan, M. Hartl, W. Chen, M. de Bono, Journal of Biological Chemistry 298 (2022).","chicago":"Artan, Murat, Markus Hartl, Weiqiang Chen, and Mario de Bono. “Depletion of Endogenously Biotinylated Carboxylases Enhances the Sensitivity of TurboID-Mediated Proximity Labeling in Caenorhabditis Elegans.” <i>Journal of Biological Chemistry</i>. Elsevier, 2022. <a href=\"https://doi.org/10.1016/j.jbc.2022.102343\">https://doi.org/10.1016/j.jbc.2022.102343</a>."},"date_created":"2022-09-11T22:01:55Z","department":[{"_id":"MaDe"}],"tmp":{"name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","image":"/images/cc_by.png","short":"CC BY (4.0)"},"month":"09","status":"public","file":[{"file_name":"2022_JBC_Artan.pdf","checksum":"e726c7b9315230e6710e0b1f1d1677e9","success":1,"file_size":2101656,"date_created":"2022-09-12T08:14:50Z","access_level":"open_access","relation":"main_file","creator":"dernst","content_type":"application/pdf","file_id":"12092","date_updated":"2022-09-12T08:14:50Z"}],"has_accepted_license":"1","acknowledgement":"We thank de Bono laboratory members for helpful comments on the article and the Mass Spec Facilities at IST Austria and Max Perutz Labs for invaluable discussions and comments on how to optimize mass spec analyses of worm samples. We are grateful to Ekaterina Lashmanova for designing the degron knock-in constructs and preparing the injection mixes for CRISPR/Cas9-mediated genome editing. All LC–MS/MS analyses were performed on instruments of the Vienna BioCenter Core Facilities instrument pool.\r\nThis work was supported by a Wellcome Investigator Award (grant no.: 209504/Z/17/Z ) to M.d.B. and an ISTplus Fellowship to M.A. (Marie Sklodowska-Curie agreement no.: 754411).","external_id":{"pmid":["35933017"],"isi":["000884241800011"]},"author":[{"last_name":"Artan","first_name":"Murat","id":"C407B586-6052-11E9-B3AE-7006E6697425","orcid":"0000-0001-8945-6992","full_name":"Artan, Murat"},{"last_name":"Hartl","first_name":"Markus","full_name":"Hartl, Markus"},{"full_name":"Chen, Weiqiang","last_name":"Chen","first_name":"Weiqiang"},{"last_name":"De Bono","first_name":"Mario","id":"4E3FF80E-F248-11E8-B48F-1D18A9856A87","full_name":"De Bono, Mario","orcid":"0000-0001-8347-0443"}],"pmid":1,"isi":1,"user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","oa_version":"Published Version"},{"ddc":["612"],"scopus_import":"1","date_updated":"2025-04-14T07:43:46Z","type":"journal_article","publication":"Journal of Biological Chemistry","article_type":"original","article_processing_charge":"Yes","abstract":[{"lang":"eng","text":"Proximity labeling provides a powerful in vivo tool to characterize the proteome of subcellular structures and the interactome of specific proteins. The nematode Caenorhabditis elegans is one of the most intensely studied organisms in biology, offering many advantages for biochemistry. Using the highly active biotin ligase TurboID, we optimize here a proximity labeling protocol for C. elegans. An advantage of TurboID is that biotin's high affinity for streptavidin means biotin-labeled proteins can be affinity-purified under harsh denaturing conditions. By combining extensive sonication with aggressive denaturation using SDS and urea, we achieved near-complete solubilization of worm proteins. We then used this protocol to characterize the proteomes of the worm gut, muscle, skin, and nervous system. Neurons are among the smallest C. elegans cells. To probe the method's sensitivity, we expressed TurboID exclusively in the two AFD neurons and showed that the protocol could identify known and previously unknown proteins expressed selectively in AFD. The active zones of synapses are composed of a protein matrix that is difficult to solubilize and purify. To test if our protocol could solubilize active zone proteins, we knocked TurboID into the endogenous elks-1 gene, which encodes a presynaptic active zone protein. We identified many known ELKS-1-interacting active zone proteins, as well as previously uncharacterized synaptic proteins. Versatile vectors and the inherent advantages of using C. elegans, including fast growth and the ability to rapidly make and functionally test knock-ins, make proximity labeling a valuable addition to the armory of this model organism."}],"language":[{"iso":"eng"}],"article_number":"101094","title":"Interactome analysis of Caenorhabditis elegans synapses by TurboID-based proximity labeling","volume":297,"year":"2021","day":"01","oa":1,"publication_status":"published","doi":"10.1016/J.JBC.2021.101094","ec_funded":1,"file_date_updated":"2021-10-11T12:20:58Z","publication_identifier":{"issn":["0021-9258"],"eissn":["1083-351X"]},"intvolume":"       297","issue":"3","_id":"10117","date_published":"2021-09-01T00:00:00Z","project":[{"grant_number":"754411","call_identifier":"H2020","name":"ISTplus - Postdoctoral Fellowships","_id":"260C2330-B435-11E9-9278-68D0E5697425"}],"quality_controlled":"1","tmp":{"name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","image":"/images/cc_by.png","short":"CC BY (4.0)"},"status":"public","month":"09","publisher":"Elsevier","citation":{"short":"M. Artan, S. Barratt, S.M. Flynn, F. Begum, M. Skehel, A. Nicolas, M. de Bono, Journal of Biological Chemistry 297 (2021).","mla":"Artan, Murat, et al. “Interactome Analysis of Caenorhabditis Elegans Synapses by TurboID-Based Proximity Labeling.” <i>Journal of Biological Chemistry</i>, vol. 297, no. 3, 101094, Elsevier, 2021, doi:<a href=\"https://doi.org/10.1016/J.JBC.2021.101094\">10.1016/J.JBC.2021.101094</a>.","ista":"Artan M, Barratt S, Flynn SM, Begum F, Skehel M, Nicolas A, de Bono M. 2021. Interactome analysis of Caenorhabditis elegans synapses by TurboID-based proximity labeling. Journal of Biological Chemistry. 297(3), 101094.","chicago":"Artan, Murat, Stephen Barratt, Sean M. Flynn, Farida Begum, Mark Skehel, Armel Nicolas, and Mario de Bono. “Interactome Analysis of Caenorhabditis Elegans Synapses by TurboID-Based Proximity Labeling.” <i>Journal of Biological Chemistry</i>. Elsevier, 2021. <a href=\"https://doi.org/10.1016/J.JBC.2021.101094\">https://doi.org/10.1016/J.JBC.2021.101094</a>.","ieee":"M. Artan <i>et al.</i>, “Interactome analysis of Caenorhabditis elegans synapses by TurboID-based proximity labeling,” <i>Journal of Biological Chemistry</i>, vol. 297, no. 3. Elsevier, 2021.","ama":"Artan M, Barratt S, Flynn SM, et al. Interactome analysis of Caenorhabditis elegans synapses by TurboID-based proximity labeling. <i>Journal of Biological Chemistry</i>. 2021;297(3). doi:<a href=\"https://doi.org/10.1016/J.JBC.2021.101094\">10.1016/J.JBC.2021.101094</a>","apa":"Artan, M., Barratt, S., Flynn, S. M., Begum, F., Skehel, M., Nicolas, A., &#38; de Bono, M. (2021). Interactome analysis of Caenorhabditis elegans synapses by TurboID-based proximity labeling. <i>Journal of Biological Chemistry</i>. Elsevier. <a href=\"https://doi.org/10.1016/J.JBC.2021.101094\">https://doi.org/10.1016/J.JBC.2021.101094</a>"},"date_created":"2021-10-10T22:01:23Z","department":[{"_id":"MaDe"},{"_id":"LifeSc"}],"acknowledgement":"We thank de Bono lab members for helpful comments on the manuscript, IST Austria and University of Vienna Mass Spec Facilities for invaluable discussions and comments for the optimization of mass spec analyses of worm samples. The biotin auxotropic E. coli strain MG1655bioB:kan was gift from John Cronan (University of Illinois) and was kindly sent to us by Jessica Feldman and Ariana Sanchez (Stanford University). dg398 pEntryslot2_mNeongreen::3XFLAG::stop and dg397 pEntryslot3_mNeongreen::3XFLAG::stop::unc-54 3′UTR entry vector were kindly shared by Dr Dominique Glauser (University of Fribourg). Codon-optimized mScarlet vector was a generous gift from Dr Manuel Zimmer (University of Vienna).","has_accepted_license":"1","file":[{"file_name":"2021_JBC_Artan.pdf","file_size":1680010,"success":1,"checksum":"19e39d36c5b9387c6dc0e89c9ae856ab","access_level":"open_access","relation":"main_file","date_created":"2021-10-11T12:20:58Z","content_type":"application/pdf","creator":"cchlebak","file_id":"10121","date_updated":"2021-10-11T12:20:58Z"}],"oa_version":"Published Version","user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","author":[{"id":"C407B586-6052-11E9-B3AE-7006E6697425","orcid":"0000-0001-8945-6992","full_name":"Artan, Murat","first_name":"Murat","last_name":"Artan"},{"first_name":"Stephen","last_name":"Barratt","id":"57740d2b-2a88-11ec-97cf-d9e6d1b39677","full_name":"Barratt, Stephen"},{"full_name":"Flynn, Sean M.","first_name":"Sean M.","last_name":"Flynn"},{"full_name":"Begum, Farida","last_name":"Begum","first_name":"Farida"},{"full_name":"Skehel, Mark","last_name":"Skehel","first_name":"Mark"},{"first_name":"Armel","last_name":"Nicolas","full_name":"Nicolas, Armel","id":"2A103192-F248-11E8-B48F-1D18A9856A87"},{"full_name":"De Bono, Mario","id":"4E3FF80E-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0001-8347-0443","last_name":"De Bono","first_name":"Mario"}],"external_id":{"isi":["000706409200006"]},"isi":1},{"publication_status":"published","doi":"10.1074/jbc.RA120.012628","main_file_link":[{"url":"https://escholarship.umassmed.edu/oapubs/4187","open_access":"1"}],"abstract":[{"lang":"eng","text":"Following its evoked release, dopamine (DA) signaling is rapidly terminated by presynaptic reuptake, mediated by the cocaine-sensitive DA transporter (DAT). DAT surface availability is dynamically regulated by endocytic trafficking, and direct protein kinase C (PKC) activation acutely diminishes DAT surface expression by accelerating DAT internalization. Previous cell line studies demonstrated that PKC-stimulated DAT endocytosis requires both Ack1 inactivation, which releases a DAT-specific endocytic brake, and the neuronal GTPase, Rit2, which binds DAT. However, it is unknown whether Rit2 is required for PKC-stimulated DAT endocytosis in DAergic terminals or whether there are region- and/or sex-dependent differences in PKC-stimulated DAT trafficking. Moreover, the mechanisms by which Rit2 controls PKC-stimulated DAT endocytosis are unknown. Here, we directly examined these important questions. Ex vivo studies revealed that PKC activation acutely decreased DAT surface expression selectively in ventral, but not dorsal, striatum. AAV-mediated, conditional Rit2 knockdown in DAergic neurons impacted baseline DAT surface:intracellular distribution in DAergic terminals from female ventral, but not dorsal, striatum. Further, Rit2 was required for PKC-stimulated DAT internalization in both male and female ventral striatum. FRET and surface pulldown studies in cell lines revealed that PKC activation drives DAT-Rit2 surface dissociation and that the DAT N terminus is required for both PKC-mediated DAT-Rit2 dissociation and DAT internalization. Finally, we found that Rit2 and Ack1 independently converge on DAT to facilitate PKC-stimulated DAT endocytosis. Together, our data provide greater insight into mechanisms that mediate PKC-regulated DAT internalization and reveal unexpected region-specific differences in PKC-stimulated DAT trafficking in bona fide DAergic terminals. "}],"language":[{"iso":"eng"}],"volume":295,"title":"Dopamine transporter trafficking and Rit2 GTPase: Mechanism of action and in vivo impact","year":"2020","day":"17","oa":1,"article_type":"original","article_processing_charge":"No","scopus_import":"1","date_updated":"2025-07-10T11:54:48Z","publication":"Journal of Biological Chemistry","type":"journal_article","oa_version":"Submitted Version","user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","pmid":1,"author":[{"full_name":"Fagan, Rita R.","last_name":"Fagan","first_name":"Rita R."},{"full_name":"Kearney, Patrick J.","first_name":"Patrick J.","last_name":"Kearney"},{"full_name":"Sweeney, Carolyn G.","first_name":"Carolyn G.","last_name":"Sweeney"},{"last_name":"Luethi","first_name":"Dino","full_name":"Luethi, Dino"},{"full_name":"Schoot Uiterkamp, Florianne E","id":"3526230C-F248-11E8-B48F-1D18A9856A87","first_name":"Florianne E","last_name":"Schoot Uiterkamp"},{"full_name":"Schicker, Klaus","first_name":"Klaus","last_name":"Schicker"},{"full_name":"Alejandro, Brian S.","first_name":"Brian S.","last_name":"Alejandro"},{"full_name":"O'Connor, Lauren C.","last_name":"O'Connor","first_name":"Lauren C."},{"full_name":"Sitte, Harald H.","last_name":"Sitte","first_name":"Harald H."},{"full_name":"Melikian, Haley E.","last_name":"Melikian","first_name":"Haley E."}],"external_id":{"pmid":["32132171"],"isi":["000530288000006"]},"isi":1,"page":"5229-5244","status":"public","month":"04","citation":{"ieee":"R. R. Fagan <i>et al.</i>, “Dopamine transporter trafficking and Rit2 GTPase: Mechanism of action and in vivo impact,” <i>Journal of Biological Chemistry</i>, vol. 295, no. 16. ASBMB Publications, pp. 5229–5244, 2020.","ama":"Fagan RR, Kearney PJ, Sweeney CG, et al. Dopamine transporter trafficking and Rit2 GTPase: Mechanism of action and in vivo impact. <i>Journal of Biological Chemistry</i>. 2020;295(16):5229-5244. doi:<a href=\"https://doi.org/10.1074/jbc.RA120.012628\">10.1074/jbc.RA120.012628</a>","apa":"Fagan, R. R., Kearney, P. J., Sweeney, C. G., Luethi, D., Schoot Uiterkamp, F. E., Schicker, K., … Melikian, H. E. (2020). Dopamine transporter trafficking and Rit2 GTPase: Mechanism of action and in vivo impact. <i>Journal of Biological Chemistry</i>. ASBMB Publications. <a href=\"https://doi.org/10.1074/jbc.RA120.012628\">https://doi.org/10.1074/jbc.RA120.012628</a>","short":"R.R. Fagan, P.J. Kearney, C.G. Sweeney, D. Luethi, F.E. Schoot Uiterkamp, K. Schicker, B.S. Alejandro, L.C. O’Connor, H.H. Sitte, H.E. Melikian, Journal of Biological Chemistry 295 (2020) 5229–5244.","mla":"Fagan, Rita R., et al. “Dopamine Transporter Trafficking and Rit2 GTPase: Mechanism of Action and in Vivo Impact.” <i>Journal of Biological Chemistry</i>, vol. 295, no. 16, ASBMB Publications, 2020, pp. 5229–44, doi:<a href=\"https://doi.org/10.1074/jbc.RA120.012628\">10.1074/jbc.RA120.012628</a>.","ista":"Fagan RR, Kearney PJ, Sweeney CG, Luethi D, Schoot Uiterkamp FE, Schicker K, Alejandro BS, O’Connor LC, Sitte HH, Melikian HE. 2020. Dopamine transporter trafficking and Rit2 GTPase: Mechanism of action and in vivo impact. Journal of Biological Chemistry. 295(16), 5229–5244.","chicago":"Fagan, Rita R., Patrick J. Kearney, Carolyn G. Sweeney, Dino Luethi, Florianne E Schoot Uiterkamp, Klaus Schicker, Brian S. Alejandro, Lauren C. O’Connor, Harald H. Sitte, and Haley E. Melikian. “Dopamine Transporter Trafficking and Rit2 GTPase: Mechanism of Action and in Vivo Impact.” <i>Journal of Biological Chemistry</i>. ASBMB Publications, 2020. <a href=\"https://doi.org/10.1074/jbc.RA120.012628\">https://doi.org/10.1074/jbc.RA120.012628</a>."},"publisher":"ASBMB Publications","date_created":"2020-05-24T22:00:59Z","department":[{"_id":"SaSi"}],"publication_identifier":{"eissn":["1083-351X"],"issn":["0021-9258"]},"issue":"16","intvolume":"       295","_id":"7880","date_published":"2020-04-17T00:00:00Z","quality_controlled":"1"},{"oa":1,"extern":"1","day":"14","year":"1994","volume":269,"title":"Brain-derived neurotrophic factor increases neurotrophin-3 expression in cerebellar granule neurons","publist_id":"1941","language":[{"iso":"eng"}],"abstract":[{"lang":"eng","text":"Neurotrophin-3 (NT-3) is a member of the neurotrophin gene family and is highly expressed in the developing rat cerebellum. Here we show that brain-derived neurotrophic factor (BDNF) increased by approximately 10-fold the NT-3 mRNA levels in cultured cerebellar granule neurons isolated from postnatal rats, whereas nerve growth factor (NGF) and NT-3 itself had no effect. The effect of BDNF was additive to that of triiodothyronine (T3), which also increased NT-3 mRNA in these neurons. The drug K252a inhibited the BDNF-mediated stimulation of NT-3 expression, suggesting an involvement of trkB receptors. Nuclear run-on experiments showed that BDNF enhanced NT-3 transcription, whereas the stability of NT-3 mRNA remained unchanged. The data presented are the first demonstration that one neurotrophin regulates the expression of another and provide evidence that NT-3 production in granule neurons is regulated by both BDNF and T3."}],"main_file_link":[{"url":"https://www.sciencedirect.com/science/article/pii/S0021925817421867?via%3Dihub","open_access":"1"}],"doi":"10.1016/s0021-9258(17)42186-7","publication_status":"published","publication":"Journal of Biological Chemistry","type":"journal_article","date_updated":"2022-06-02T10:23:48Z","scopus_import":"1","article_processing_charge":"No","article_type":"original","acknowledgement":"We thank Dorothea Stratmann and Karin Angermayer for skillful technical assistance.","author":[{"full_name":"Leingärtner, Axel","first_name":"Axel","last_name":"Leingärtner"},{"first_name":"Carl-Philipp J","last_name":"Heisenberg","id":"39427864-F248-11E8-B48F-1D18A9856A87","full_name":"Heisenberg, Carl-Philipp J","orcid":"0000-0002-0912-4566"},{"full_name":"Kolbeck, Roland","first_name":"Roland","last_name":"Kolbeck"},{"full_name":"Thoenen, Hans","last_name":"Thoenen","first_name":"Hans"},{"full_name":"Lindholm, Dan","last_name":"Lindholm","first_name":"Dan"}],"user_id":"ea97e931-d5af-11eb-85d4-e6957dddbf17","oa_version":"None","quality_controlled":"1","date_published":"1994-01-14T00:00:00Z","_id":"4179","publication_identifier":{"issn":["0021-9258"],"eissn":["1083-351X"]},"issue":"2","intvolume":"       269","date_created":"2018-12-11T12:07:25Z","citation":{"ama":"Leingärtner A, Heisenberg C-PJ, Kolbeck R, Thoenen H, Lindholm D. Brain-derived neurotrophic factor increases neurotrophin-3 expression in cerebellar granule neurons. <i>Journal of Biological Chemistry</i>. 1994;269(2):828-830. doi:<a href=\"https://doi.org/10.1016/s0021-9258(17)42186-7\">10.1016/s0021-9258(17)42186-7</a>","apa":"Leingärtner, A., Heisenberg, C.-P. J., Kolbeck, R., Thoenen, H., &#38; Lindholm, D. (1994). Brain-derived neurotrophic factor increases neurotrophin-3 expression in cerebellar granule neurons. <i>Journal of Biological Chemistry</i>. American Society for Biochemistry and Molecular Biology. <a href=\"https://doi.org/10.1016/s0021-9258(17)42186-7\">https://doi.org/10.1016/s0021-9258(17)42186-7</a>","ieee":"A. Leingärtner, C.-P. J. Heisenberg, R. Kolbeck, H. Thoenen, and D. Lindholm, “Brain-derived neurotrophic factor increases neurotrophin-3 expression in cerebellar granule neurons,” <i>Journal of Biological Chemistry</i>, vol. 269, no. 2. American Society for Biochemistry and Molecular Biology, pp. 828–830, 1994.","chicago":"Leingärtner, Axel, Carl-Philipp J Heisenberg, Roland Kolbeck, Hans Thoenen, and Dan Lindholm. “Brain-Derived Neurotrophic Factor Increases Neurotrophin-3 Expression in Cerebellar Granule Neurons.” <i>Journal of Biological Chemistry</i>. American Society for Biochemistry and Molecular Biology, 1994. <a href=\"https://doi.org/10.1016/s0021-9258(17)42186-7\">https://doi.org/10.1016/s0021-9258(17)42186-7</a>.","short":"A. Leingärtner, C.-P.J. Heisenberg, R. Kolbeck, H. Thoenen, D. Lindholm, Journal of Biological Chemistry 269 (1994) 828–830.","mla":"Leingärtner, Axel, et al. “Brain-Derived Neurotrophic Factor Increases Neurotrophin-3 Expression in Cerebellar Granule Neurons.” <i>Journal of Biological Chemistry</i>, vol. 269, no. 2, American Society for Biochemistry and Molecular Biology, 1994, pp. 828–30, doi:<a href=\"https://doi.org/10.1016/s0021-9258(17)42186-7\">10.1016/s0021-9258(17)42186-7</a>.","ista":"Leingärtner A, Heisenberg C-PJ, Kolbeck R, Thoenen H, Lindholm D. 1994. Brain-derived neurotrophic factor increases neurotrophin-3 expression in cerebellar granule neurons. Journal of Biological Chemistry. 269(2), 828–830."},"publisher":"American Society for Biochemistry and Molecular Biology","month":"01","status":"public","page":"828 - 830"},{"oa_version":"Published Version","user_id":"ea97e931-d5af-11eb-85d4-e6957dddbf17","author":[{"id":"499F3ABC-F248-11E8-B48F-1D18A9856A87","full_name":"Shigemoto, Ryuichi","orcid":"0000-0001-8761-9444","last_name":"Shigemoto","first_name":"Ryuichi"},{"last_name":"Yokota","first_name":"Yoshifumi","full_name":"Yokota, Yoshifumi"},{"full_name":"Tsuchida, Kunihiro","first_name":"Kunihiro","last_name":"Tsuchida"},{"full_name":"Nakanishi, Shigetada","last_name":"Nakanishi","first_name":"Shigetada"}],"pmid":1,"external_id":{"pmid":["2153106 "]},"acknowledgement":"This work was supported in part by research grants from the Ministry Education, Science and Culture of Japan; the Institute of Physical and Chemical Research; and the Science and Technology Agency of Japan. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 USC. Section 1734 solely to indicate this fact.","page":"623 - 628","status":"public","month":"01","citation":{"mla":"Shigemoto, Ryuichi, et al. “Cloning and Expression of a Rat Neuromedin K Receptor CDNA.” <i>Journal of Biological Chemistry</i>, vol. 265, no. 2, American Society for Biochemistry and Molecular Biology, 1990, pp. 623–28, doi:<a href=\"https://doi.org/10.1016/s0021-9258(19)40095-1 \">10.1016/s0021-9258(19)40095-1 </a>.","short":"R. Shigemoto, Y. Yokota, K. Tsuchida, S. Nakanishi, Journal of Biological Chemistry 265 (1990) 623–628.","ista":"Shigemoto R, Yokota Y, Tsuchida K, Nakanishi S. 1990. Cloning and expression of a rat neuromedin K receptor cDNA. Journal of Biological Chemistry. 265(2), 623–628.","chicago":"Shigemoto, Ryuichi, Yoshifumi Yokota, Kunihiro Tsuchida, and Shigetada Nakanishi. “Cloning and Expression of a Rat Neuromedin K Receptor CDNA.” <i>Journal of Biological Chemistry</i>. American Society for Biochemistry and Molecular Biology, 1990. <a href=\"https://doi.org/10.1016/s0021-9258(19)40095-1 \">https://doi.org/10.1016/s0021-9258(19)40095-1 </a>.","apa":"Shigemoto, R., Yokota, Y., Tsuchida, K., &#38; Nakanishi, S. (1990). Cloning and expression of a rat neuromedin K receptor cDNA. <i>Journal of Biological Chemistry</i>. American Society for Biochemistry and Molecular Biology. <a href=\"https://doi.org/10.1016/s0021-9258(19)40095-1 \">https://doi.org/10.1016/s0021-9258(19)40095-1 </a>","ama":"Shigemoto R, Yokota Y, Tsuchida K, Nakanishi S. Cloning and expression of a rat neuromedin K receptor cDNA. <i>Journal of Biological Chemistry</i>. 1990;265(2):623-628. doi:<a href=\"https://doi.org/10.1016/s0021-9258(19)40095-1 \">10.1016/s0021-9258(19)40095-1 </a>","ieee":"R. Shigemoto, Y. Yokota, K. Tsuchida, and S. Nakanishi, “Cloning and expression of a rat neuromedin K receptor cDNA,” <i>Journal of Biological Chemistry</i>, vol. 265, no. 2. American Society for Biochemistry and Molecular Biology, pp. 623–628, 1990."},"publisher":"American Society for Biochemistry and Molecular Biology","date_created":"2018-12-11T11:57:55Z","issue":"2","publication_identifier":{"eissn":["1083-351X"],"issn":["0021-9258"]},"intvolume":"       265","_id":"2480","date_published":"1990-01-15T00:00:00Z","quality_controlled":"1","publication_status":"published","doi":"10.1016/s0021-9258(19)40095-1 ","main_file_link":[{"open_access":"1","url":"https://www.sciencedirect.com/science/article/pii/S0021925819400951"}],"abstract":[{"lang":"eng","text":"Functional cDNA clones for rat neuromedin K receptor were isolated from a rat brain cDNA library by cross-hybridization with the bovine substance K recepor cDNA. Injection of the mRNA synthesized in vitro from the cloned cDNA into Xenopus oocytes elicited electrophysiological responses to tachykinins, with the most potent sensitivity being to neuromedin K. Ligand-binding displacement in membranes of mammalian COS cells transfected with the cDNA indicated the rank order of affinity of the receptor to tachykinins; neuromedin K &gt; substance K &gt; substance P. The hybridization analysis showed that the neuromedin K receptor mRNA is expressed in both the brain and the peripheral tissues at different levels. The rat neuromedin K receptor consists of 452 amino acid residues and belongs to the family of G protein-coupled receptors, which are thought to have seven transmembrane domains. The sequence comparison of the rat neuromedin K, substance P, and substance K receptors revealed that these receptors are highly conserved in the seven transmembrane domains and the cytoplasmic sides of the receptors. They also show some structural characteristics, including the common presence of histidine residues in transmembrane segments V and VI and the difference in the numbers and distributions of serine and threonine residues as possible phosphorylation sites in the cytoplasmic regions. This paper thus presents the first comprehensive analysis of the molecular nature of the multiple peptide receptors that exhibit similar but pharmacologically distinguishable activities."}],"language":[{"iso":"eng"}],"publist_id":"4421","volume":265,"title":"Cloning and expression of a rat neuromedin K receptor cDNA","year":"1990","day":"15","extern":"1","oa":1,"article_type":"original","article_processing_charge":"No","scopus_import":"1","date_updated":"2022-02-24T11:07:05Z","type":"journal_article","publication":"Journal of Biological Chemistry"},{"date_updated":"2025-06-26T09:18:02Z","type":"journal_article","publication":"Journal of Biological Chemistry","scopus_import":"1","article_processing_charge":"No","article_type":"original","day":"25","extern":"1","oa":1,"volume":264,"title":"Molecular characterization of a functional cDNA for rat substance P receptor","year":"1989","language":[{"iso":"eng"}],"publist_id":"4374","abstract":[{"lang":"eng","text":"This paper describes the amino acid sequence of the rat substance P receptor and its comparison with that of the rat substance K receptor on the basis of molecular cloning and sequence analysis. From a rat brain cDNA library constructed with an RNA expression vector, we identified a cDNA mixture containing a functional substance P receptor cDNA by examining electrophysiologically a receptor expression following injection of the mRNAs synthesized in vitro into Xenopus oocytes. A receptor cDNA clone was then isolated by cross-hybridization with the bovine substance K receptor DNA. The clone was confirmed by selective binding of substance P to the cloned receptor expressed in mammalian COS cells. The deduced amino acid sequence (407 amino acid residues) possesses seven putative membrane spanning domains and shows a sequence similarity to the members of G-protein-coupled receptors. The rat substance P and substance K receptor are very similar in both size and amino acid sequences, particularly in the putative transmembrane similarity is in marked contrast to the sequence divergence in the amino- and carboxyl-terminal regions and the third cytoplasmic loop. The observed sequence similarytity and divergence would thus contribute to the expression of similar but pharmacological regions and the first and second cytoplasmic loops. This distinguishable activities of the two tachykinin receptors."}],"main_file_link":[{"open_access":"1","url":"https://doi.org/10.1016/S0021-9258(19)84619-7"}],"doi":"10.1016/S0021-9258(19)84619-7","publication_status":"published","quality_controlled":"1","date_published":"1989-10-25T00:00:00Z","issue":"30","publication_identifier":{"eissn":["1083-351X"],"issn":["0021-9258"]},"intvolume":"       264","_id":"2525","date_created":"2018-12-11T11:58:11Z","citation":{"ista":"Yokota Y, Sasai Y, Tanaka K, Fujiwara T, Tsuchida K, Shigemoto R, Kakizuka A, Ohkubo H, Nakanishi S. 1989. Molecular characterization of a functional cDNA for rat substance P receptor. Journal of Biological Chemistry. 264(30), 17649–17652.","short":"Y. Yokota, Y. Sasai, K. Tanaka, T. Fujiwara, K. Tsuchida, R. Shigemoto, A. Kakizuka, H. Ohkubo, S. Nakanishi, Journal of Biological Chemistry 264 (1989) 17649–17652.","mla":"Yokota, Yoshifumi, et al. “Molecular Characterization of a Functional CDNA for Rat Substance P Receptor.” <i>Journal of Biological Chemistry</i>, vol. 264, no. 30, American Society for Biochemistry and Molecular Biology, 1989, pp. 17649–52, doi:<a href=\"https://doi.org/10.1016/S0021-9258(19)84619-7\">10.1016/S0021-9258(19)84619-7</a>.","chicago":"Yokota, Yoshifumi, Yoshiki Sasai, Kohichi Tanaka, Tsutomu Fujiwara, Kunihiro Tsuchida, Ryuichi Shigemoto, Akira Kakizuka, Hiroaki Ohkubo, and Shigetada Nakanishi. “Molecular Characterization of a Functional CDNA for Rat Substance P Receptor.” <i>Journal of Biological Chemistry</i>. American Society for Biochemistry and Molecular Biology, 1989. <a href=\"https://doi.org/10.1016/S0021-9258(19)84619-7\">https://doi.org/10.1016/S0021-9258(19)84619-7</a>.","ieee":"Y. Yokota <i>et al.</i>, “Molecular characterization of a functional cDNA for rat substance P receptor,” <i>Journal of Biological Chemistry</i>, vol. 264, no. 30. American Society for Biochemistry and Molecular Biology, pp. 17649–17652, 1989.","ama":"Yokota Y, Sasai Y, Tanaka K, et al. Molecular characterization of a functional cDNA for rat substance P receptor. <i>Journal of Biological Chemistry</i>. 1989;264(30):17649-17652. doi:<a href=\"https://doi.org/10.1016/S0021-9258(19)84619-7\">10.1016/S0021-9258(19)84619-7</a>","apa":"Yokota, Y., Sasai, Y., Tanaka, K., Fujiwara, T., Tsuchida, K., Shigemoto, R., … Nakanishi, S. (1989). Molecular characterization of a functional cDNA for rat substance P receptor. <i>Journal of Biological Chemistry</i>. American Society for Biochemistry and Molecular Biology. <a href=\"https://doi.org/10.1016/S0021-9258(19)84619-7\">https://doi.org/10.1016/S0021-9258(19)84619-7</a>"},"publisher":"American Society for Biochemistry and Molecular Biology","status":"public","month":"10","page":"17649 - 17652","acknowledgement":"This work was supported in part by research grants from the Ministry of Education, Science and Culture of Japan, the Institute of Physical and Chemical Research, and the Science and Technology Agency of Japan. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. ","pmid":1,"author":[{"first_name":"Yoshifumi","last_name":"Yokota","full_name":"Yokota, Yoshifumi"},{"first_name":"Yoshiki","last_name":"Sasai","full_name":"Sasai, Yoshiki"},{"full_name":"Tanaka, Kohichi","last_name":"Tanaka","first_name":"Kohichi"},{"full_name":"Fujiwara, Tsutomu","first_name":"Tsutomu","last_name":"Fujiwara"},{"full_name":"Tsuchida, Kunihiro","last_name":"Tsuchida","first_name":"Kunihiro"},{"orcid":"0000-0001-8761-9444","full_name":"Shigemoto, Ryuichi","id":"499F3ABC-F248-11E8-B48F-1D18A9856A87","first_name":"Ryuichi","last_name":"Shigemoto"},{"full_name":"Kakizuka, Akira","first_name":"Akira","last_name":"Kakizuka"},{"full_name":"Ohkubo, Hiroaki","first_name":"Hiroaki","last_name":"Ohkubo"},{"full_name":"Nakanishi, Shigetada","first_name":"Shigetada","last_name":"Nakanishi"}],"external_id":{"pmid":["2478537"]},"oa_version":"Published Version","user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87"}]