{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,5]],"date-time":"2026-09-05T01:23:11Z","timestamp":1788571391374,"version":"build-2803163510"},"reference-count":58,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2023,9,8]],"date-time":"2023-09-08T00:00:00Z","timestamp":1694131200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2023,9,8]],"date-time":"2023-09-08T00:00:00Z","timestamp":1694131200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["T32 HL120822"],"award-info":[{"award-number":["T32 HL120822"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R35 GM131821"],"award-info":[{"award-number":["R35 GM131821"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01 GM125082"],"award-info":[{"award-number":["R01 GM125082"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Inositol depletion is a hypothesized mechanism of action of mood stabilization drugs used in the treatment of bipolar disorder. It was previously reported that the mood stabilizer valproate (VPA) increased phosphorylation of <jats:italic>myo<\/jats:italic>-inositol-3-phosphate synthases (MIPS), the rate limiting enzyme of inositol synthesis. Phosphosites were identified and examination of site-directed mutants suggested that phosphorylation leads to decreased enzymatic activity. In this study, we examined the extent of MIPS phosphorylation in response to VPA and used two interaction screens to identify protein kinases that interact with MIPS. Using an epitope tagged MIPS construct, we determined the fraction of phosphorylated MIPS to be very low (less than 2% of total), and we could not detect phosphorylation of untagged MIPS in response to VPA. In vitro analyses of phosphorylation revealed that putative protein kinases, PKC and CKII, have low specificity toward MIPS. These findings suggest that VPA likely depletes inositol via a mechanism other than MIPS phosphorylation. Consistent with this, mRNA levels of the MIPS-encoding gene <jats:italic>INO1<\/jats:italic> and MIPS protein levels were significantly reduced during the mid-log growth phase in response to VPA treatment. These findings suggest that the mechanism whereby VPA causes inositol depletion is by reducing expression of the rate-limiting enzyme MIPS.<\/jats:p>","DOI":"10.1038\/s41598-023-41936-2","type":"journal-article","created":{"date-parts":[[2023,9,8]],"date-time":"2023-09-08T12:02:47Z","timestamp":1694174567000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Valproate regulates inositol synthesis by reducing expression of myo-inositol-3-phosphate synthase"],"prefix":"10.1038","volume":"13","author":[{"given":"Kendall C.","family":"Case","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rachel J.","family":"Beltman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mary Kay H.","family":"Pflum","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miriam L.","family":"Greenberg","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2023,9,8]]},"reference":[{"key":"41936_CR1","doi-asserted-by":"crossref","unstructured":"Case, K. C., Salsaa, M., Yu, W. & Greenberg, M. L. in Lipid Signaling in Human diseases regulation of inositol biosynthesis: Balancing health and pathophysiology, 221\u2013260 (Springer, 2018).","DOI":"10.1007\/164_2018_181"},{"key":"41936_CR2","doi-asserted-by":"publisher","first-page":"5079","DOI":"10.3390\/molecules25215079","volume":"25","author":"S Chatree","year":"2020","unstructured":"Chatree, S., Thongmaen, N., Tantivejkul, K., Sitticharoon, C. & Vucenik, I. Role of inositols and inositol phosphates in energy metabolism. Molecules 25, 5079. https:\/\/doi.org\/10.3390\/molecules25215079 (2020).","journal-title":"Molecules"},{"key":"41936_CR3","doi-asserted-by":"publisher","first-page":"10437","DOI":"10.1074\/jbc.M116.714816","volume":"291","author":"W Yu","year":"2016","unstructured":"Yu, W., Ye, C. & Greenberg, M. L. Inositol hexakisphosphate kinase 1 (IP6K1) regulates inositol synthesis in mammalian cells. J. Biol. Chem. 291, 10437\u201310444. https:\/\/doi.org\/10.1074\/jbc.M116.714816 (2016).","journal-title":"J. Biol. Chem."},{"key":"41936_CR4","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbc.2022.102363","author":"P Lazcano","year":"2022","unstructured":"Lazcano, P., Schmidtke, M. W., Onu, C. & Greenberg, M. L. Phosphatidic acid inhibits inositol synthesis by inducing nuclear translocation of kinase IP6K1 and repression of myo-inositol-3-P synthase. J. Biol. Chem. https:\/\/doi.org\/10.1016\/j.jbc.2022.102363 (2022).","journal-title":"J. Biol. Chem."},{"key":"41936_CR5","doi-asserted-by":"publisher","first-page":"249","DOI":"10.1007\/s10555-022-10032-8","volume":"41","author":"KC Case","year":"2022","unstructured":"Case, K. C., Schmidtke, M. W. & Greenberg, M. L. The paradoxical role of inositol in cancer: A consequence of the metabolic state of a tumor. Cancer Metastasis Rev. 41, 249\u2013254. https:\/\/doi.org\/10.1007\/s10555-022-10032-8 (2022).","journal-title":"Cancer Metastasis Rev."},{"key":"41936_CR6","doi-asserted-by":"publisher","first-page":"369","DOI":"10.1042\/BJ20130118","volume":"452","author":"MS Wilson","year":"2013","unstructured":"Wilson, M. S., Livermore, T. M. & Saiardi, A. Inositol pyrophosphates: Between signalling and metabolism. Biochem. J. 452, 369\u2013379. https:\/\/doi.org\/10.1042\/BJ20130118 (2013).","journal-title":"Biochem. J."},{"key":"41936_CR7","doi-asserted-by":"publisher","first-page":"159","DOI":"10.1042\/BJ20140361","volume":"461","author":"PG Billcliff","year":"2014","unstructured":"Billcliff, P. G. & Lowe, M. Inositol lipid phosphatases in membrane trafficking and human disease. Biochem. J. 461, 159\u2013175. https:\/\/doi.org\/10.1042\/BJ20140361 (2014).","journal-title":"Biochem. J."},{"key":"41936_CR8","doi-asserted-by":"publisher","first-page":"651","DOI":"10.1038\/nature05185","volume":"443","author":"G Di Paolo","year":"2006","unstructured":"Di Paolo, G. & De Camilli, P. Phosphoinositides in cell regulation and membrane dynamics. Nature 443, 651\u2013657. https:\/\/doi.org\/10.1038\/nature05185 (2006).","journal-title":"Nature"},{"key":"41936_CR9","doi-asserted-by":"publisher","first-page":"477","DOI":"10.1007\/s00441-014-1806-z","volume":"357","author":"MJ Berridge","year":"2014","unstructured":"Berridge, M. J. Calcium signalling and psychiatric disease: Bipolar disorder and schizophrenia. Cell Tissue Res 357, 477\u2013492. https:\/\/doi.org\/10.1007\/s00441-014-1806-z (2014).","journal-title":"Cell Tissue Res"},{"key":"41936_CR10","doi-asserted-by":"publisher","first-page":"403","DOI":"10.3390\/medicina55080403","volume":"55","author":"P Dome","year":"2019","unstructured":"Dome, P., Rihmer, Z. & Gonda, X. Suicide risk in bipolar disorder: A brief review. Medicina 55, 403 (2019).","journal-title":"Medicina"},{"key":"41936_CR11","doi-asserted-by":"publisher","first-page":"1761","DOI":"10.1016\/j.pnpbp.2008.08.012","volume":"32","author":"F Marmol","year":"2008","unstructured":"Marmol, F. Lithium: bipolar disorder and neurodegenerative diseases possible cellular mechanisms of the therapeutic effects of lithium. Prog. Neuropsychopharmacol. Biol. Psychiatry 32, 1761\u20131771 (2008).","journal-title":"Prog. Neuropsychopharmacol. Biol. Psychiatry"},{"key":"41936_CR12","doi-asserted-by":"publisher","first-page":"131","DOI":"10.4103\/0019-5545.111449","volume":"55","author":"J Trivedi","year":"2013","unstructured":"Trivedi, J., Sareen, H., Yadav, V. S. & Rai, S. B. Prescription pattern of mood stabilizers for bipolar disorder at a tertiary health care centre in north India. Indian J. Psychiatry 55, 131 (2013).","journal-title":"Indian J. Psychiatry"},{"key":"41936_CR13","doi-asserted-by":"publisher","first-page":"587","DOI":"10.1042\/bj2060587","volume":"206","author":"MJ Berridge","year":"1982","unstructured":"Berridge, M. J., Downes, C. P. & Hanley, M. R. Lithium amplifies agonist-dependent phosphatidylinositol responses in brain and salivary glands. Biochem. J. 206, 587\u2013595. https:\/\/doi.org\/10.1042\/bj2060587 (1982).","journal-title":"Biochem. J."},{"key":"41936_CR14","doi-asserted-by":"publisher","first-page":"197","DOI":"10.1038\/341197a0","volume":"341","author":"MJ Berridge","year":"1989","unstructured":"Berridge, M. J. & Irvine, R. F. Inositol phosphates and cell signalling. Nature 341, 197\u2013205. https:\/\/doi.org\/10.1038\/341197a0 (1989).","journal-title":"Nature"},{"key":"41936_CR15","doi-asserted-by":"publisher","first-page":"10896","DOI":"10.1016\/S0021-9258(19)70391-3","volume":"255","author":"LM Hallcher","year":"1980","unstructured":"Hallcher, L. M. & Sherman, W. R. The effects of lithium ion and other agents on the activity of myo-inositol-1-phosphatase from bovine brain. J. Biol. Chem. 255, 10896\u201310901 (1980).","journal-title":"J. Biol. Chem."},{"key":"41936_CR16","doi-asserted-by":"publisher","first-page":"277","DOI":"10.1016\/0893-133X(94)00088-H","volume":"12","author":"R Vadnal","year":"1995","unstructured":"Vadnal, R. & Parthasarathy, R. Myo-inositol monophosphatase: diverse effects of lithium, carbamazepine, and valproate. Neuropsychopharmacology 12, 277\u2013285. https:\/\/doi.org\/10.1016\/0893-133X(94)00088-H (1995).","journal-title":"Neuropsychopharmacology"},{"key":"41936_CR17","doi-asserted-by":"publisher","first-page":"5309","DOI":"10.1111\/febs.13070","volume":"281","author":"A Dutta","year":"2014","unstructured":"Dutta, A., Bhattacharyya, S., Dutta, D. & Das, A. K. Structural elucidation of the binding site and mode of inhibition of Li+ and Mg2+ in inositol monophosphatase. FEBS J. 281, 5309\u20135324 (2014).","journal-title":"FEBS J."},{"key":"41936_CR18","doi-asserted-by":"publisher","first-page":"2398","DOI":"10.1021\/ci300172r","volume":"52","author":"S Lu","year":"2012","unstructured":"Lu, S. et al. Insights into the role of magnesium triad in myo-inositol monophosphatase: Metal mechanism, substrate binding, and lithium therapy. J. Chem. Inf. Model. 52, 2398\u20132409 (2012).","journal-title":"J. Chem. Inf. Model."},{"key":"41936_CR19","doi-asserted-by":"publisher","first-page":"26822","DOI":"10.1074\/jbc.M113.479121","volume":"288","author":"RM Deranieh","year":"2013","unstructured":"Deranieh, R. M., He, Q., Caruso, J. A. & Greenberg, M. L. Phosphorylation regulates myo-inositol-3-phosphate synthase: A novel regulatory mechanism of inositol biosynthesis. J. Biol. Chem. 288, 26822\u201326833. https:\/\/doi.org\/10.1074\/jbc.M113.479121 (2013).","journal-title":"J. Biol. Chem."},{"key":"41936_CR20","doi-asserted-by":"publisher","first-page":"1595","DOI":"10.1046\/j.1365-2958.2003.03641.x","volume":"49","author":"S Ju","year":"2003","unstructured":"Ju, S. & Greenberg, M. L. Valproate disrupts regulation of inositol responsive genes and alters regulation of phospholipid biosynthesis. Mol. Microbiol. 49, 1595\u20131603. https:\/\/doi.org\/10.1046\/j.1365-2958.2003.03641.x (2003).","journal-title":"Mol. Microbiol."},{"key":"41936_CR21","doi-asserted-by":"publisher","first-page":"84","DOI":"10.1016\/S0006-8993(00)02797-9","volume":"880","author":"T O\u2019Donnell","year":"2000","unstructured":"O\u2019Donnell, T. et al. Chronic lithium and sodium valproate both decrease the concentration of myo-inositol and increase the concentration of inositol monophosphates in rat brain. Brain Res. 880, 84\u201391 (2000).","journal-title":"Brain Res."},{"key":"41936_CR22","doi-asserted-by":"publisher","first-page":"868","DOI":"10.1016\/j.biopsych.2004.08.027","volume":"56","author":"G Shaltiel","year":"2004","unstructured":"Shaltiel, G. et al. Valproate decreases inositol biosynthesis. Biol. Psychiatry 56, 868\u2013874. https:\/\/doi.org\/10.1016\/j.biopsych.2004.08.027 (2004).","journal-title":"Biol. Psychiatry"},{"key":"41936_CR23","doi-asserted-by":"publisher","first-page":"15466","DOI":"10.1074\/jbc.M004179200","volume":"276","author":"DL Vaden","year":"2001","unstructured":"Vaden, D. L., Ding, D., Peterson, B. & Greenberg, M. L. Lithium and valproate decrease inositol mass and increase expression of the yeast INO1 and INO2 genes for inositol biosynthesis. J. Biol. Chem. 276, 15466\u201315471. https:\/\/doi.org\/10.1074\/jbc.M004179200 (2001).","journal-title":"J. Biol. Chem."},{"key":"41936_CR24","doi-asserted-by":"publisher","first-page":"127","DOI":"10.15252\/emmm.201404137","volume":"7","author":"M Schiebler","year":"2015","unstructured":"Schiebler, M. et al. Functional drug screening reveals anticonvulsants as enhancers of mTOR-independent autophagic killing of Mycobacterium tuberculosis through inositol depletion. EMBO Mol. Med. 7, 127\u2013139. https:\/\/doi.org\/10.15252\/emmm.201404137 (2015).","journal-title":"EMBO Mol. Med."},{"key":"41936_CR25","doi-asserted-by":"publisher","first-page":"135","DOI":"10.2217\/fnl-2016-0003","volume":"11","author":"W Yu","year":"2016","unstructured":"Yu, W. & Greenberg, M. L. Inositol depletion, GSK3 inhibition and bipolar disorder. Future Neurol. 11, 135\u2013148. https:\/\/doi.org\/10.2217\/fnl-2016-0003 (2016).","journal-title":"Future Neurol."},{"key":"41936_CR26","doi-asserted-by":"publisher","first-page":"1099","DOI":"10.1042\/BST0371099","volume":"37","author":"RM Deranieh","year":"2009","unstructured":"Deranieh, R. M. & Greenberg, M. L. Cellular consequences of inositol depletion. Biochem. Soc. Trans. 37, 1099\u20131103. https:\/\/doi.org\/10.1042\/BST0371099 (2009).","journal-title":"Biochem. Soc. Trans."},{"key":"41936_CR27","doi-asserted-by":"publisher","first-page":"0182534","DOI":"10.1371\/journal.pone.0182534","volume":"12","author":"W Yu","year":"2017","unstructured":"Yu, W., Daniel, J., Mehta, D., Maddipati, K. R. & Greenberg, M. L. MCK1 is a novel regulator of myo-inositol phosphate synthase (MIPS) that is required for inhibition of inositol synthesis by the mood stabilizer valproate. PLoS ONE 12, 0182534. https:\/\/doi.org\/10.1371\/journal.pone.0182534 (2017).","journal-title":"PLoS ONE"},{"key":"41936_CR28","doi-asserted-by":"publisher","first-page":"1188","DOI":"10.1091\/mbc.E15-12-0844","volume":"27","author":"DI Kim","year":"2016","unstructured":"Kim, D. I. et al. An improved smaller biotin ligase for BioID proximity labeling. Mol. Biol. Cell 27, 1188\u20131196. https:\/\/doi.org\/10.1091\/mbc.E15-12-0844 (2016).","journal-title":"Mol. Biol. Cell"},{"key":"41936_CR29","doi-asserted-by":"publisher","first-page":"2199","DOI":"10.1074\/mcp.M116.066654","volume":"16","author":"N Opitz","year":"2017","unstructured":"Opitz, N. et al. Capturing the Asc1p\/receptor for activated C kinase 1 (RACK1) microenvironment at the head region of the 40S ribosome with quantitative BioID in yeast. Mol. Cell. Proteomics 16, 2199\u20132218 (2017).","journal-title":"Mol. Cell. Proteomics"},{"key":"41936_CR30","doi-asserted-by":"publisher","first-page":"11","DOI":"10.1002\/0471140864.ps1923s74","volume":"74","author":"KJ Roux","year":"2013","unstructured":"Roux, K. J., Kim, D. I. & Burke, B. BioID: a screen for protein-protein interactions. Curr. Protoc. Protein Sci. 74, 11\u201314. https:\/\/doi.org\/10.1002\/0471140864.ps1923s74 (2013).","journal-title":"Curr. Protoc. Protein Sci."},{"key":"41936_CR31","doi-asserted-by":"publisher","first-page":"e539","DOI":"10.1002\/cpz1.539","volume":"2","author":"RJ Beltman","year":"2022","unstructured":"Beltman, R. J. & Pflum, M. K. H. Kinase-catalyzed crosslinking and immunoprecipitation (K-CLIP) to explore kinase-substrate pairs. Curr. Protoc. 2, e539 (2022).","journal-title":"Curr. Protoc."},{"key":"41936_CR32","doi-asserted-by":"publisher","first-page":"16299","DOI":"10.1021\/jacs.8b10160","volume":"140","author":"S Garre","year":"2018","unstructured":"Garre, S., Gamage, A. K., Faner, T. R., Dedigama-Arachchige, P. & Pflum, M. K. H. Identification of kinases and interactors of p53 using kinase-catalyzed cross-linking and immunoprecipitation. J. Am. Chem. Soc. 140, 16299\u201316310 (2018).","journal-title":"J. Am. Chem. Soc."},{"key":"41936_CR33","doi-asserted-by":"publisher","first-page":"1367","DOI":"10.1038\/nbt.1511","volume":"26","author":"J Cox","year":"2008","unstructured":"Cox, J. & Mann, M. MaxQuant enables high peptide identification rates, individualized ppb-range mass accuracies and proteome-wide protein quantification. Nat. Biotechnol. 26, 1367\u20131372 (2008).","journal-title":"Nat. Biotechnol."},{"key":"41936_CR34","doi-asserted-by":"publisher","first-page":"14370","DOI":"10.1096\/fj.201901712R","volume":"33","author":"H Bao","year":"2019","unstructured":"Bao, H. et al. Lithium targeting of AMPK protects against cisplatin-induced acute kidney injury by enhancing autophagy in renal proximal tubular epithelial cells. FASEB J. 33, 14370\u201314381. https:\/\/doi.org\/10.1096\/fj.201901712R (2019).","journal-title":"FASEB J."},{"key":"41936_CR35","doi-asserted-by":"publisher","first-page":"158552","DOI":"10.1016\/j.bbalip.2019.158552","volume":"1865","author":"Y Lee","year":"2020","unstructured":"Lee, Y. et al. Lithium chloride promotes lipid accumulation through increased reactive oxygen species generation. Biochim. Biophys. Acta Mol. Cell Biol. Lipids 1865, 158552 (2020).","journal-title":"Biochim. Biophys. Acta Mol. Cell Biol. Lipids"},{"key":"41936_CR36","doi-asserted-by":"publisher","first-page":"101110","DOI":"10.1016\/j.jbc.2021.101110","volume":"297","author":"M Salsaa","year":"2021","unstructured":"Salsaa, M. et al. Valproate activates the Snf1 kinase in Saccharomyces cerevisiae by decreasing the cytosolic pH. J. Biol. Chem. 297, 101110. https:\/\/doi.org\/10.1016\/j.jbc.2021.101110 (2021).","journal-title":"J. Biol. Chem."},{"key":"41936_CR37","doi-asserted-by":"publisher","first-page":"3803","DOI":"10.1016\/j.molcel.2021.08.025","volume":"81","author":"CC Hsu","year":"2021","unstructured":"Hsu, C. C. et al. Inositol serves as a natural inhibitor of mitochondrial fission by directly targeting AMPK. Mol. Cell 81, 3803\u20133819. https:\/\/doi.org\/10.1016\/j.molcel.2021.08.025 (2021).","journal-title":"Mol. Cell"},{"key":"41936_CR38","doi-asserted-by":"publisher","first-page":"3635","DOI":"10.1093\/nar\/gkg584","volume":"31","author":"JC Obenauer","year":"2003","unstructured":"Obenauer, J. C., Cantley, L. C. & Yaffe, M. B. Scansite 2.0: Proteome-wide prediction of cell signaling interactions using short sequence motifs. Nucleic Acids Res. 31, 3635\u20133641 (2003).","journal-title":"Nucleic Acids Res."},{"key":"41936_CR39","doi-asserted-by":"publisher","first-page":"1633","DOI":"10.1002\/pmic.200300771","volume":"4","author":"N Blom","year":"2004","unstructured":"Blom, N., Sicheritz-Ponten, T., Gupta, R., Gammeltoft, S. & Brunak, S. Prediction of post-translational glycosylation and phosphorylation of proteins from the amino acid sequence. Proteomics 4, 1633\u20131649. https:\/\/doi.org\/10.1002\/pmic.200300771 (2004).","journal-title":"Proteomics"},{"key":"41936_CR40","doi-asserted-by":"publisher","first-page":"11290","DOI":"10.1074\/jbc.M112.346023","volume":"287","author":"H-S Choi","year":"2012","unstructured":"Choi, H.-S. et al. Pho85p-Pho80p phosphorylation of Yeast Pah1p phosphatidate phosphatase regulates its activity, location, abundance, and function in lipid metabolism. J. Biol. Chem. 287, 11290\u201311301 (2012).","journal-title":"J. Biol. Chem."},{"key":"41936_CR41","doi-asserted-by":"publisher","first-page":"18818","DOI":"10.1074\/jbc.M114.581462","volume":"289","author":"WM Su","year":"2014","unstructured":"Su, W. M., Han, G. S. & Carman, G. M. Cross-talk phosphorylations by protein kinase C and Pho85p-Pho80p protein kinase regulate Pah1p phosphatidate phosphatase abundance in Saccharomyces cerevisiae. J. Biol. Chem 289, 18818\u201318830. https:\/\/doi.org\/10.1074\/jbc.M114.581462 (2014).","journal-title":"J. Biol. Chem"},{"key":"41936_CR42","doi-asserted-by":"publisher","first-page":"33364","DOI":"10.1074\/jbc.M112.402339","volume":"287","author":"W-M Su","year":"2012","unstructured":"Su, W.-M., Han, G.-S., Casciano, J. & Carman, G. M. Protein kinase A-mediated phosphorylation of Pah1p phosphatidate phosphatase functions in conjunction with the Pho85p-Pho80p and Cdc28p-cyclin B kinases to regulate lipid synthesis in yeast. J. Biol. Chem. 287, 33364\u201333376 (2012).","journal-title":"J. Biol. Chem."},{"key":"41936_CR43","doi-asserted-by":"publisher","first-page":"9974","DOI":"10.1074\/jbc.M116.726588","volume":"291","author":"LS Hsieh","year":"2016","unstructured":"Hsieh, L. S., Su, W. M., Han, G. S. & Carman, G. M. Phosphorylation of yeast Pah1 phosphatidate phosphatase by casein kinase II regulates its function in lipid metabolism. J. Biol. Chem. 291, 9974\u20139990. https:\/\/doi.org\/10.1074\/jbc.M116.726588 (2016).","journal-title":"J. Biol. Chem."},{"key":"41936_CR44","doi-asserted-by":"publisher","first-page":"83","DOI":"10.1007\/s11010-013-1597-7","volume":"378","author":"RN Parthasarathy","year":"2013","unstructured":"Parthasarathy, R. N. et al. Rat brain myo-inositol 3-phosphate synthase is a phosphoprotein. Mol. Cell Biochem. 378, 83\u201389. https:\/\/doi.org\/10.1007\/s11010-013-1597-7 (2013).","journal-title":"Mol. Cell Biochem."},{"key":"41936_CR45","doi-asserted-by":"publisher","first-page":"43089","DOI":"10.1074\/jbc.M305956200","volume":"278","author":"FM Vaz","year":"2003","unstructured":"Vaz, F. M., Houtkooper, R. H., Valianpour, F., Barth, P. G. & Wanders, R. J. Only one splice variant of the human TAZ gene encodes a functional protein with a role in cardiolipin metabolism. J. Biol. Chem. 278, 43089\u201343094. https:\/\/doi.org\/10.1074\/jbc.M305956200 (2003).","journal-title":"J. Biol. Chem."},{"key":"41936_CR46","doi-asserted-by":"crossref","unstructured":"Becker, D. M. & Guarente, L. in Methods in enzymology Vol. 194 [12] High-efficiency transformation of yeast by electroporation, 182\u2013187 (Elsevier, 1991).","DOI":"10.1016\/0076-6879(91)94015-5"},{"key":"41936_CR47","doi-asserted-by":"publisher","first-page":"745","DOI":"10.1002\/yea.1628","volume":"25","author":"M Orlova","year":"2008","unstructured":"Orlova, M., Barrett, L. & Kuchin, S. Detection of endogenous Snf1 and its activation state: Application to Saccharomyces and Candida species. Yeast 25, 745\u2013754 (2008).","journal-title":"Yeast"},{"key":"41936_CR48","doi-asserted-by":"crossref","unstructured":"Agrawal, G. K. & Thelen, J. J. in The protein protocols handbook A modified Pro-Q Diamond staining protocol for phosphoprotein detection in polyacrylamide gels, 579\u2013585 (Springer, 2009).","DOI":"10.1007\/978-1-59745-198-7_54"},{"key":"41936_CR49","doi-asserted-by":"publisher","first-page":"376","DOI":"10.2144\/03352pt01","volume":"35","author":"RC Krieg","year":"2003","unstructured":"Krieg, R. C., Paweletz, C. P., Liotta, L. A. & Petricoin, E. F. 3rd. Dilution of protein gel stain results in retention of staining capacity. Biotechniques 35, 376\u2013378. https:\/\/doi.org\/10.2144\/03352pt01 (2003).","journal-title":"Biotechniques"},{"key":"41936_CR50","doi-asserted-by":"publisher","first-page":"e64","DOI":"10.1002\/cpps.64","volume":"93","author":"Z Nagy","year":"2018","unstructured":"Nagy, Z., Comer, S. & Smolenski, A. Analysis of protein phosphorylation using Phos-Tag gels. Curr. Protoc. Protein Sci. 93, e64 (2018).","journal-title":"Curr. Protoc. Protein Sci."},{"key":"41936_CR51","doi-asserted-by":"publisher","first-page":"6649","DOI":"10.1038\/s41598-019-43039-3","volume":"9","author":"J Xu","year":"2019","unstructured":"Xu, J. et al. A fixation method for the optimisation of western blotting. Sci. Rep. 9, 6649 (2019).","journal-title":"Sci. Rep."},{"key":"41936_CR52","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/1477-5956-9-45","volume":"9","author":"MM Sousa","year":"2011","unstructured":"Sousa, M. M., Steen, K. W., Hagen, L. & Slupphaug, G. Antibody cross-linking and target elution protocols used for immunoprecipitation significantly modulate signal-to noise ratio in downstream 2D-PAGE analysis. Proteome Sci. 9, 1\u20138 (2011).","journal-title":"Proteome Sci."},{"key":"41936_CR53","doi-asserted-by":"publisher","first-page":"1522","DOI":"10.1016\/j.bbrc.2017.09.168","volume":"493","author":"JS Cheah","year":"2017","unstructured":"Cheah, J. S. & Yamada, S. A simple elution strategy for biotinylated proteins bound to streptavidin conjugated beads using excess biotin and heat. Biochem. Biophys. Res. Commun. 493, 1522\u20131527 (2017).","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"41936_CR54","doi-asserted-by":"publisher","first-page":"522","DOI":"10.1194\/jlr.M022798","volume":"53","author":"Z Xu","year":"2012","unstructured":"Xu, Z., Su, W. M. & Carman, G. M. Fluorescence spectroscopy measures yeast PAH1-encoded phosphatidate phosphatase interaction with liposome membranes. J. Lipid Res. 53, 522\u2013528. https:\/\/doi.org\/10.1194\/jlr.M022798 (2012).","journal-title":"J. Lipid Res."},{"key":"41936_CR55","doi-asserted-by":"publisher","first-page":"9210","DOI":"10.1074\/jbc.M600425200","volume":"281","author":"G-S Han","year":"2006","unstructured":"Han, G.-S., Wu, W.-I. & Carman, G. M. The saccharomyces cerevisiae lipin homolog is a Mg2+-dependent phosphatidate phosphatase enzyme*\u2666. J. Biol. Chem. 281, 9210\u20139218 (2006).","journal-title":"J. Biol. Chem."},{"key":"41936_CR56","doi-asserted-by":"publisher","first-page":"34699","DOI":"10.1074\/jbc.M114.614883","volume":"289","author":"W-M Su","year":"2014","unstructured":"Su, W.-M., Han, G.-S. & Carman, G. M. Yeast Nem1-Spo7 protein phosphatase activity on Pah1 phosphatidate phosphatase is specific for the Pho85-Pho80 protein kinase phosphorylation sites. J. Biol. Chem. 289, 34699\u201334708 (2014).","journal-title":"J. Biol. Chem."},{"key":"41936_CR57","doi-asserted-by":"publisher","first-page":"997","DOI":"10.1006\/jmbi.2000.4417","volume":"306","author":"DA Jeffery","year":"2001","unstructured":"Jeffery, D. A., Springer, M., King, D. S. & O\u2019Shea, E. K. Multi-site phosphorylation of Pho4 by the cyclin-CDK Pho80-Pho85 is semi-processive with site preference. J. Mol. Biol. 306, 997\u20131010 (2001).","journal-title":"J. Mol. Biol."},{"key":"41936_CR58","doi-asserted-by":"publisher","first-page":"102221","DOI":"10.1016\/j.jbc.2022.102221","volume":"298","author":"S Khondker","year":"2022","unstructured":"Khondker, S., Kwiatek, J. M., Han, G.-S. & Carman, G. M. Glycogen synthase kinase homolog Rim11 regulates lipid synthesis through the phosphorylation of Pah1 phosphatidate phosphatase in yeast. J. Biol. Chem. 298, 102221 (2022).","journal-title":"J. Biol. Chem."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-023-41936-2.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-023-41936-2","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-023-41936-2.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,11,19]],"date-time":"2023-11-19T02:09:17Z","timestamp":1700359757000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-023-41936-2"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,9,8]]},"references-count":58,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2023,12]]}},"alternative-id":["41936"],"URL":"https:\/\/doi.org\/10.1038\/s41598-023-41936-2","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,9,8]]},"assertion":[{"value":"25 March 2023","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"4 September 2023","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"8 September 2023","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"14844"}}