Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

BCAA–BCKA axis regulates WAT browning through acetylation of PRDM16

Nature Metabolism volume 4, pages 106–122 (2022)Cite this article

Subjects

Abstract

The link between branched-chain amino acids (BCAAs) and obesity has been known for decades but the functional role of BCAA metabolism in white adipose tissue (WAT) of obese individuals remains vague. Here, we show that mice with adipose tissue knockout of Bcat2, which converts BCAAs to branched-chain keto acids (BCKAs), are resistant to high-fat diet-induced obesity due to increased inguinal WAT browning and thermogenesis. Mechanistically, acetyl-CoA derived from BCKA suppresses WAT browning by acetylation of PR domain-containing protein 16 (PRDM16) at K915, disrupting the interaction between PRDM16 and peroxisome proliferator-activated receptor-γ (PPARγ) to maintain WAT characteristics. Depletion of BCKA-derived acetyl-CoA robustly prompts WAT browning and energy expenditure. In contrast, BCKA supplementation re-establishes high-fat diet-induced obesity in Bcat2 knockout mice. Moreover, telmisartan, an anti-hypertension drug, significantly represses Bcat2 activity via direct binding, resulting in enhanced WAT browning and reduced adiposity. Strikingly, BCKA supplementation reverses the lean phenotype conferred by telmisartan. Thus, we uncover the critical role of the BCAA–BCKA axis in WAT browning.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: Loss of Bcat2 in adipocytes protects mice against obesity.
Fig. 2: Loss of Bcat2 in adipose tissue promotes WAT browning.
Fig. 3: BCAA-derived acetyl-CoA disrupts binding of Prdm16 to Pparγ.
Fig. 4: BCKA re-establishes HFD-induced obesity in Bcat2 KO mice.
Fig. 5: TEL inhibits BCAT2 activity by direct binding.
Fig. 6: TEL protects mice against obesity via inhibiting Bcat2.
Fig. 7: BCKA reversed the lean phenotype conferred by TEL.

Similar content being viewed by others

Data availability

The acetylation sites were found in the protein PTM database (www.phosphosite.org). The structure of BCAT2 was downloaded from the Protein Data Bank (https://www.rcsb.org/structure/1kta). The structure of telmisartan was downloaded from PubChem (https://pubchem.ncbi.nlm.nih.gov/compound/65999). The data used to generate the main results shown in the main figures and extended figures are available as source data. Source data including uncropped western blots are provided with this paper or upon request from the corresponding authors. All data supporting the findings of this study are available from the corresponding authors on reasonable request.

References

  1. Felig, P., Marliss, E. & Cahill, G. F. Jr. Plasma amino acid levels and insulin secretion in obesity. N. Engl. J. Med. 281, 811–816 (1969).

    Article  CAS  PubMed  Google Scholar 

  2. Lynch, C. J. & Adams, S. H. Branched-chain amino acids in metabolic signalling and insulin resistance. Nat. Rev. Endocrinol. 10, 723–736 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Cirulli, E. T. et al. Profound perturbation of the metabolome in obesity is associated with health risk. Cell Metab. 29, 488–500 e482 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Wang, T. J. et al. Metabolite profiles and the risk of developing diabetes. Nat. Med. 17, 448–453 (2011).

    Article  PubMed  PubMed Central  Google Scholar 

  5. Newgard, C. B. et al. A branched-chain amino acid-related metabolic signature that differentiates obese and lean humans and contributes to insulin resistance. Cell Metab. 9, 311–326 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Green, C. R. et al. Branched-chain amino acid catabolism fuels adipocyte differentiation and lipogenesis. Nat. Chem. Biol. 12, 15–21 (2016).

    Article  CAS  PubMed  Google Scholar 

  7. Wallace, M. et al. Enzyme promiscuity drives branched-chain fatty acid synthesis in adipose tissues. Nat. Chem. Biol. 14, 1021–1031 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Neinast, M. D. et al. Quantitative analysis of the whole-body metabolic fate of branched-chain amino acids. Cell Metab. 29, 417–429 e414 (2019).

    Article  CAS  PubMed  Google Scholar 

  9. Solon-Biet, S. M. et al. Branched chain amino acids impact health and lifespan indirectly via amino acid balance and appetite control. Nat. Metab. 1, 532–545 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Cheng, Y. et al. Leucine deprivation decreases fat mass by stimulation of lipolysis in white adipose tissue and upregulation of uncoupling protein 1 (UCP1) in brown adipose tissue. Diabetes 59, 17–25 (2010).

    Article  CAS  PubMed  Google Scholar 

  11. White, P. J. et al. Branched-chain amino acid restriction in Zucker-fatty rats improves muscle insulin sensitivity by enhancing efficiency of fatty acid oxidation and acyl-glycine export. Mol. Metab. 5, 538–551 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Fontana, L. et al. Decreased consumption of branched-chain amino acids improves metabolic health. Cell Rep. 16, 520–530 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Cummings, N. E. et al. Restoration of metabolic health by decreased consumption of branched-chain amino acids. J. Physiol. 596, 623–645 (2018).

    Article  CAS  PubMed  Google Scholar 

  14. Wu, Y. et al. Very-low-protein diets lead to reduced food intake and weight loss, linked to inhibition of hypothalamic mTOR signaling, in mice. Cell Metab. 33, 888–904 e886 (2021).

    Article  CAS  PubMed  Google Scholar 

  15. Yu, D. et al. The adverse metabolic effects of branched-chain amino acids are mediated by isoleucine and valine. Cell Metab. 33, 905–922 e906 (2021).

    Article  CAS  PubMed  Google Scholar 

  16. Chuang, D. T., Ku, L. S. & Cox, R. P. Thiamin-responsive maple-syrup-urine disease: decreased affinity of the mutant branched-chain alpha-keto acid dehydrogenase for alpha-ketoisovalerate and thiamin pyrophosphate. Proc. Natl Acad. Sci. USA 79, 3300–3304 (1982).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Peinemann, F. & Danner, D. J. Maple syrup urine disease 1954 to 1993. J. Inherit. Metab. Dis. 17, 3–15 (1994).

    Article  CAS  PubMed  Google Scholar 

  18. Zinnanti, W. J. et al. Dual mechanism of brain injury and novel treatment strategy in maple syrup urine disease. Brain 132, 903–918 (2009).

    Article  PubMed  PubMed Central  Google Scholar 

  19. Lu, G. et al. Protein phosphatase 2Cm is a critical regulator of branched-chain amino acid catabolism in mice and cultured cells. J. Clin. Invest. 119, 1678–1687 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. She, P. et al. Disruption of BCATm in mice leads to increased energy expenditure associated with the activation of a futile protein turnover cycle. Cell Metab. 6, 181–194 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Yoneshiro, T. et al. BCAA catabolism in brown fat controls energy homeostasis through SLC25A44. Nature 572, 614–619 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Bluher, M. Obesity: global epidemiology and pathogenesis. Nat. Rev. Endocrinol. 15, 288–298 (2019).

    Article  PubMed  Google Scholar 

  23. van Baak, M. A. & Mariman, E. C. M. Mechanisms of weight regain after weight loss—the role of adipose tissue. Nat. Rev. Endocrinol. 15, 274–287 (2019).

    Article  PubMed  Google Scholar 

  24. Morigny, P., Boucher, J., Arner, P. & Langin, D. Lipid and glucose metabolism in white adipocytes: pathways, dysfunction and therapeutics. Nat. Rev. Endocrinol. 17, 276–295 (2021).

    Article  CAS  PubMed  Google Scholar 

  25. White, P. J. & Newgard, C. B. Branched-chain amino acids in disease. Science 363, 582–583 (2019).

    Article  CAS  PubMed  Google Scholar 

  26. Neinast, M., Murashige, D. & Arany, Z. Branched chain amino acids. Annu. Rev. Physiol. 81, 139–164 (2019).

    Article  CAS  PubMed  Google Scholar 

  27. Eguchi, J. et al. Transcriptional control of adipose lipid handling by IRF4. Cell Metab. 13, 249–259 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Jimenez, V. et al. In vivo adeno-associated viral vector-mediated genetic engineering of white and brown adipose tissue in adult mice. Diabetes 62, 4012–4022 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Liu, G. Y. & Sabatini, D. M. mTOR at the nexus of nutrition, growth, ageing and disease. Nat. Rev. Mol. Cell Biol. 21, 183–203 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Mina, A. I. et al. CalR: a web-based analysis tool for indirect calorimetry experiments. Cell Metab. 28, 656–666 e651 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Frontini, A. & Cinti, S. Distribution and development of brown adipocytes in the murine and human adipose organ. Cell Metab. 11, 253–256 (2010).

    Article  CAS  PubMed  Google Scholar 

  32. Chouchani, E. T., Kazak, L. & Spiegelman, B. M. New advances in adaptive thermogenesis: UCP1 and beyond. Cell Metab. 29, 27–37 (2019).

    Article  CAS  PubMed  Google Scholar 

  33. Balaz, M. et al. Inhibition of mevalonate pathway prevents adipocyte browning in mice and men by affecting protein prenylation. Cell Metab. 29, 901–916 e908 (2019).

    Article  CAS  PubMed  Google Scholar 

  34. Shimizu, I. et al. Vascular rarefaction mediates whitening of brown fat in obesity. J. Clin. Invest. 124, 2099–2112 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Kuipers, E. N. et al. A single day of high-fat diet feeding induces lipid accumulation and insulin resistance in brown adipose tissue in mice. Am. J. Physiol. Endocrinol. Metab. 317, E820–E830 (2019).

    Article  CAS  PubMed  Google Scholar 

  36. van Marken Lichtenbelt, W. D. et al. Cold-activated brown adipose tissue in healthy men. N. Engl. J. Med. 360, 1500–1508 (2009).

    Article  PubMed  Google Scholar 

  37. Shimizu, I. & Walsh, K. The whitening of brown fat and its implications for weight management in obesity. Curr. Obes. Rep. 4, 224–229 (2015).

    Article  PubMed  Google Scholar 

  38. Becher, T. et al. Brown adipose tissue is associated with cardiometabolic health. Nat. Med. 27, 58–65 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Seale, P. et al. Prdm16 determines the thermogenic program of subcutaneous white adipose tissue in mice. J. Clin. Invest. 121, 96–105 (2011).

    Article  CAS  PubMed  Google Scholar 

  40. Seale, P. et al. Transcriptional control of brown fat determination by PRDM16. Cell Metab. 6, 38–54 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Kissig, M., Shapira, S. N. & Seale, P. SnapShot: brown and beige adipose thermogenesis. Cell 166, 258–258 e251 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Wang, W. & Seale, P. Control of brown and beige fat development. Nat. Rev. Mol. Cell Biol. 17, 691–702 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Lin, J., Handschin, C. & Spiegelman, B. M. Metabolic control through the PGC-1 family of transcription coactivators. Cell Metab. 1, 361–370 (2005).

    Article  PubMed  Google Scholar 

  44. Son, S. M. et al. Leucine signals to mTORC1 via its metabolite acetyl-coenzyme A. Cell Metab. 29, 192–201 e197 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Carrer, A. et al. Acetyl-CoA metabolism supports multistep pancreatic tumorigenesis. Cancer Discov. 9, 416–435 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Marino, G. et al. Regulation of autophagy by cytosolic acetyl-coenzyme A. Mol. Cell 53, 710–725 (2014).

    Article  CAS  PubMed  Google Scholar 

  47. Camussi, G., Bussolino, F., Salvidio, G. & Baglioni, C. Tumor necrosis factor/cachectin stimulates peritoneal macrophages, polymorphonuclear neutrophils, and vascular endothelial cells to synthesize and release platelet-activating factor. J. Exp. Med. 166, 1390–1404 (1987).

    Article  CAS  PubMed  Google Scholar 

  48. Gao, X. et al. Acetate functions as an epigenetic metabolite to promote lipid synthesis under hypoxia. Nat. Commun. 7, 11960 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Madeo, F., Pietrocola, F., Eisenberg, T. & Kroemer, G. Caloric restriction mimetics: towards a molecular definition. Nat. Rev. Drug Discov. 13, 727–740 (2014).

    Article  CAS  PubMed  Google Scholar 

  50. Choudhary, C., Weinert, B. T., Nishida, Y., Verdin, E. & Mann, M. The growing landscape of lysine acetylation links metabolism and cell signalling. Nat. Rev. Mol. Cell Biol. 15, 536–550 (2014).

    Article  CAS  PubMed  Google Scholar 

  51. Pietrocola, F., Galluzzi, L., Bravo-San Pedro, J. M., Madeo, F. & Kroemer, G. Acetyl coenzyme A: a central metabolite and second messenger. Cell Metab. 21, 805–821 (2015).

    Article  CAS  PubMed  Google Scholar 

  52. Qian, H. et al. HDAC6-mediated acetylation of lipid droplet-binding protein CIDEC regulates fat-induced lipid storage. J. Clin. Invest. 127, 1353–1369 (2017).

    Article  PubMed  PubMed Central  Google Scholar 

  53. Yan, Y. et al. HDAC6 suppresses age-dependent ectopic fat accumulation by maintaining the proteostasis of PLIN2 in Drosophila. Dev. Cell 43, 99–111 e115 (2017).

    Article  CAS  PubMed  Google Scholar 

  54. Lieber, A. D. et al. Loss of HDAC6 alters gut microbiota and worsens obesity. FASEB J. 33, 1098–1109 (2019).

    Article  CAS  PubMed  Google Scholar 

  55. Seale, P. et al. PRDM16 controls a brown fat/skeletal muscle switch. Nature 454, 961–967 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  56. Block, K. P. & Harper, A. E. High levels of dietary amino and branched-chain ɑ-keto acids alter plasma and brain amino acid concentrations in rats. J. Nutr. 121, 663–671 (1991).

    Article  CAS  PubMed  Google Scholar 

  57. Crowell, P. L. et al. High branched-chain alpha-keto acid intake, branched-chain alpha-keto acid dehydrogenase activity, and plasma and brain amino acid and plasma keto acid concentrations in rats. Am. J. Clin. Nutr. 52, 313–319 (1990).

    Article  CAS  PubMed  Google Scholar 

  58. Cohen, P. et al. Ablation of PRDM16 and beige adipose causes metabolic dysfunction and a subcutaneous to visceral fat switch. Cell 156, 304–316 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  59. Wang, W. et al. A PRDM16-driven metabolic signal from adipocytes regulates precursor cell fate. Cell Metab. 30, 174–189.e5 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  60. Hales, C. M., Fryar, C. D., Carroll, M. D., Freedman, D. S. & Ogden, C. L. Trends in obesity and severe obesity prevalence in US youth and adults by sex and age, 2007–2008 to 2015–2016. JAMA 319, 1723–1725 (2018).

    Article  PubMed  PubMed Central  Google Scholar 

  61. Jang, C. et al. A branched-chain amino acid metabolite drives vascular fatty acid transport and causes insulin resistance. Nat. Med. 22, 421–426 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  62. Deshmukh, A. S. et al. Proteomics-based comparative mapping of the secretomes of human brown and white adipocytes reveals EPDR1 as a novel batokine. Cell Metab. 30, 963–975 e967 (2019).

    Article  CAS  PubMed  Google Scholar 

  63. She, P. et al. Obesity-related elevations in plasma leucine are associated with alterations in enzymes involved in branched-chain amino acid metabolism. Am. J. Physiol. Endocrinol. Metab. 293, E1552–E1563 (2007).

    Article  CAS  PubMed  Google Scholar 

  64. Lackey, D. E. et al. Regulation of adipose branched-chain amino acid catabolism enzyme expression and cross-adipose amino acid flux in human obesity. Am. J. Physiol. Endocrinol. Metab. 304, E1175–E1187 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  65. Shin, A. C. et al. Brain insulin lowers circulating BCAA levels by inducing hepatic BCAA catabolism. Cell Metab. 20, 898–909 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  66. White, P. J. et al. The BCKDH kinase and phosphatase integrate BCAA and lipid metabolism via regulation of ATP-citrate lyase. Cell Metab. 27, 1281–1293 e1287 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  67. Ericksen, R. E. et al. Loss of BCAA catabolism during carcinogenesis enhances mTORC1 activity and promotes tumor development and progression. Cell Metab. 29, 1151–1165 e1156 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  68. Kotzbeck, P. et al. Brown adipose tissue whitening leads to brown adipocyte death and adipose tissue inflammation. J. Lipid Res. 59, 784–794 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  69. Li, L. et al. Telmisartan improves insulin resistance of skeletal muscle through peroxisome proliferator-activated receptor-δ activation. Diabetes 62, 762–774 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  70. Sugimoto, K. et al. Telmisartan but not valsartan increases caloric expenditure and protects against weight gain and hepatic steatosis. Hypertension 47, 1003–1009 (2006).

    Article  CAS  PubMed  Google Scholar 

  71. Araki, K. et al. Telmisartan prevents obesity and increases the expression of uncoupling protein 1 in diet-induced obese mice. Hypertension 48, 51–57 (2006).

    Article  CAS  PubMed  Google Scholar 

  72. Benson, S. C. et al. Identification of telmisartan as a unique angiotensin II receptor antagonist with selective PPARγ-modulating activity. Hypertension 43, 993–1002 (2004).

    Article  CAS  PubMed  Google Scholar 

  73. Schupp, M., Janke, J., Clasen, R., Unger, T. & Kintscher, U. Angiotensin type 1 receptor blockers induce peroxisome proliferator-activated receptor-gamma activity. Circulation 109, 2054–2057 (2004).

    Article  CAS  PubMed  Google Scholar 

  74. Yki-Jarvinen, H. Thiazolidinediones. N. Engl. J. Med. 351, 1106–1118 (2004).

    Article  PubMed  Google Scholar 

  75. Murakami, K. et al. The effects of telmisartan treatment on the abdominal fat depot in patients with metabolic syndrome and essential hypertension: Abdominal fat Depot Intervention Program of Okayama (ADIPO). Diab. Vasc. Dis. Res 10, 93–96 (2013).

    Article  PubMed  Google Scholar 

  76. Kakuma, T. et al. Telmisartan reduced abdominal circumference and body weight with decreasing triglyceride level in patients with type 2 diabetes and metabolic syndrome. Obes. Res. Clin. Pract. 4, e83–e162 (2010).

    Article  PubMed  Google Scholar 

  77. Negro, R., Formoso, G. & Hassan, H. The effects of irbesartan and telmisartan on metabolic parameters and blood pressure in obese, insulin resistant, hypertensive patients. J. Endocrinol. Invest. 29, 957–961 (2006).

    Article  CAS  PubMed  Google Scholar 

  78. Priest, C. & Tontonoz, P. Inter-organ cross-talk in metabolic syndrome. Nat. Metab. 1, 1177–1188 (2019).

    Article  PubMed  Google Scholar 

  79. Galarraga, M. et al. Adiposoft: automated software for the analysis of white adipose tissue cellularity in histological sections. J. Lipid Res. 53, 2791–2796 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  80. Kidder, B. L., Hu, G. & Zhao, K. ChIP-Seq: technical considerations for obtaining high-quality data. Nat. Immunol. 12, 918–922 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  81. Basu, S. S. & Blair, I. A. SILEC: a protocol for generating and using isotopically labeled coenzyme A mass spectrometry standards. Nat. Protoc. 7, 1–12 (2011).

    Article  PubMed  PubMed Central  Google Scholar 

  82. Davoodi, J. et al. Overexpression and characterization of the human mitochondrial and cytosolic branched-chain aminotransferases. J. Biol. Chem. 273, 4982–4989 (1998).

    Article  CAS  PubMed  Google Scholar 

  83. Cooper, A. J. L., Conway, M. & Hutson, S. M. A continuous 96-well plate spectrophotometric assay for branched-chain amino acid aminotransferases. Anal. Biochem. 308, 100–105 (2002).

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank members of the Lei Laboratory for discussion throughout this study and the Biomedical Core Facility of Fudan University for technical support. We thank Q.-R. Ding for providing Adiponectin-Cre mice. This work was supported by the National Key R&D Program of China (grant nos. 2020YFA0803400/2020YFA0803402 and 2019YFA0801703 to Q.-Y.L.), the Natural Science Foundation of China (grant nos. 81790250/81790253, 91959202 and 82121004 to Q.-Y.L.; grant no. 81902821 to Q.-X.M; grant no. 81872240 to M.Y.; grant no. 81770844 and 82070870 to H.-Y.H) and the Innovation Program of Shanghai Municipal Education Commission (grant no. N173606 to Q.-Y.L.).

Author information

Authors and Affiliations

  1. Fudan University Shanghai Cancer Center & Institutes of Biomedical Sciences; Cancer Institutes; Key Laboratory of Breast Cancer in Shanghai; Shanghai Key Laboratory of Radiation Oncology; The Shanghai Key Laboratory of Medical Epigenetics; Department of Oncology; State Key Laboratory of Medical Neurobiology, Shanghai Medical College, Fudan University, Shanghai, China

    Qi-Xiang Ma, Wen-Ying Zhu, Xiao-Chen Lu, Jin-Tao Li, Miao Yin & Qun-Ying Lei

  2. Key Laboratory of Metabolism and Molecular Medicine of Chinese Ministry of Education, Department of Biochemistry and Molecular Biology of School of Basic Medical Sciences, Fudan University, Shanghai, China

    Duo Jiang & Hai-Yan Huang

  3. Institute of Molecular and Cell Biology, Agency for Science, Technology and Research (A*STAR), Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Republic of Singapore

    Feng Xu

  4. Institutes of Biomedical Sciences, Fudan University, Shanghai, China

    Lei Zhang

  5. Hongqiao International Institute of Medicine, Shanghai Tongren Hospital/Faculty of Basic Medicine, Chemical Biology Division of Shanghai Universities E-Institutes, Key Laboratory of Cell Differentiation and Apoptosis of the Chinese Ministry of Education, Shanghai Jiao Tong University School of Medicine, Shanghai, China

    Ying-Li Wu

  6. State Key Laboratory of Cell Biology, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, Shanghai, China

    Zheng-Jun Chen

Authors
  1. Qi-Xiang Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wen-Ying Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xiao-Chen Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Duo Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Feng Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jin-Tao Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Lei Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Ying-Li Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Zheng-Jun Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Miao Yin
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Hai-Yan Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Qun-Ying Lei
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Q.-X.M., W.-Y.Z., H.-Y.H., D.J. X.-C.L. and J.-T.L. performed the experiments. Q.-X.M., W.-Y.Z. and X.-C.L. analysed the data. Q.-X.M., M.Y., F.X. and H.-Y.H. cowrote the manuscript. Y.-L.W. provided the FDA-approved drug library. Z.-J.C. gave advice for animal experiments. L.Z. performed the LC–MS experiment. M.Y., H.-Y.H. and F.X. provided intellectual discussion. Q.-Y.L. conceived the idea, designed and supervised the study, analysed the data and cowrote the manuscript.

Corresponding authors

Correspondence to Miao Yin, Hai-Yan Huang or Qun-Ying Lei.

Ethics declarations

Competing interests

The authors declare no competing interests.

Peer review

Peer review information

Nature Metabolism thanks Feifan Guo and the other, anonymous, reviewers for their contribution to the peer review of this work. Isabella Samuelson was the primary handling editor.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data

Extended Data Fig. 1 Loss of Bcat2 in adipocytes protects mice against obesity.

(a) Bcat1 is not expressed in iWAT, gWAT and BAT. Bcat2 protein levels as indicated were quantified and normalized against Tubulin. Representative result from 2 independent biological experiments. (b and c) Bcat2 expression in indicated organs from adipose tissue specific Bcat2KO mice. Bcat2 protein levels as indicated were quantified and normalized against Tubulin. n=1. (d) Plasma BCAA levels in Bcat2WT and Bcat2KO mice. n=5–6 mice. (e) Body weight of Bcat2WT and Bcat2KO mice fed with normal chow diet. n=5–6 mice. (f) Food intake is similar in Bcat2WT and Bcat2KO mice fed with normal chow food. Representative mean value from 5 mice in a cage. (g) Bcat2 KO prevents HFD-induced weight gain. Body weight was measured after 13-week HFD feeding. n=6 mice. (h) Computed tomography imaging showed Bcat2 KO prevents HFD-induced fat tissue weight gain. n=3 mice. (i) High fat Food intake is similar in Bcat2WT and Bcat2KO mice. (j) GTT and ITT experiment in Bcat2WT and Bcat2KO mice. n=6 mice. (k) Bcat2 KO has no effect on other main organ weights. Weights of indicated organs derived from Bcat2WT and Bcat2KO mice fed with HFD were measured. n=6 mice. (l) Bcat2 KO prevents HFD-induced fatty liver. Representative result from 4 mice. (m) Bcat2 KO decreases adipocyte size. Adipocyte sizes of iWAT from Bcat2WT and Bcat2KO mice fed with HFD were calculated from the H&E staining results. n=6 per group. (n) No morphological change is observed in muscle from Bcat2KO mouse via HE staining analysis. Representative H&E staining of muscle derived from Bcat2WT and Bcat2KO mice fed with HFD. Representative result from 4 mice. Bars and error bars represent mean values and SDs, with biologically individual data points shown. n, cohort size. P values are derived from unpaired, two-tailed t-test.

Source data

Extended Data Fig. 2 Bcat2 in iWAT protects mice against obesity.

Loss of (a-b) Cre is expressed and Bcat2 is KO in iWAT from injecting AAV-Cre mice but not other organs or AAV-GFP mice, proved by IHC and WB. Bcat2 protein levels as indicated were quantified and normalized against Tubulin. n=2 mice. (c) iWAT-Bcat2 KO prevents HFD-induced weight gain. n=5 mice. (d) Food intake is similar in Bcat2WT and Bcat iWAT-KO mice fed with normal chow food. Representative mean value from 5 mice in a cage. (e) GTT, ITT, and serum markers indicated from WT and Bcat iWAT-KO mice. n=5 mice. (f) iWAT-Bcat2 KO prevents HFD-induced fatty liver. Representative result from 4 mice. (g) Bcat2 KO has no effect on mTORc1 activation. iWAT tissue from overnight starvation mice as a negative control and re-feed as positive control. p-4ebp protein levels as indicated were quantified and normalized against total 4ebp. n=3 mice. Bars and error bars represent mean values and SDs, with biologically individual data points shown. n, cohort size. P values are derived from unpaired, two-tailed t-test.

Source data

Extended Data Fig. 3 Metabolic changes of Bcat2KO mice.

(a) Bcat2 KO enhances whole body energy expenditure. Oxygen consumption, carbon dioxide generation, RER and heat production of mice fed with HFD were measured. n=5 mice. (b) Bcat2 KO has no effect on mice movement activity. n=5 mice. (c) Oxygen consumption, carbon dioxide generation, and heat production data were analyzed with total body mass as covariate. n=5 mice. (d) Thermal infrared imaging showed the temperature at interscapular area of Bcat2KO mice is significantly higher than that of Bcat2WT mice. n=4 mice. (e) Ucp1 and Cidea protein expression in iWAT from different group of mice. BAT was used as a positive control. Ucp1 protein levels from the indicated tissues were quantified and normalized against Tubulin. n=3 mice. (f) Bcat2 KO increases Ucp1 expression in BAT compared to WT fed with HFD. Representative images of Ucp1 immunohistochemistry (IHC) in BAT. WT from normal chow diet (NCD) as a control. Representative result from 4 mice. (g) iWAT-Bcat2 KO enhances whole body energy expenditure. Oxygen consumption, carbon dioxide generation, RER and heat production of mice fed with HFD were measured. n=5. (h) Oxygen consumption, carbon dioxide generation, and heat production data were analyzed with total body mass as covariate. n=5. (i) iWAT-Bcat2 KO has no effect on mice movement activity. n=5. (j) Thermal infrared imaging showed the temperature at interscapular area of Bcat2 iWAT-KO mice is significantly higher than that of Bcat2WT mice. n=4. (k) iWAT-Bcat2 KO increases Ucp1 expression in BAT compared to WT fed with HFD. Representative result from 4 mice. (l) High BCAA hyper-activates mTORc1 and disturbs Ucp1 expression. Ucp1 protein levels from the indicated tissues were quantified and normalized against Tubulin. Representative result from 2 independent biological experiments. Bars and error bars represent mean values and SDs, with biologically individual data points shown. P values are derived from unpaired, two-tailed t-test (except for c and h). Energy expenditure in c and h was analyzed by ANCOVA, with body weight included as a covariat.

Source data

Extended Data Fig. 4 Mechanism of browning in Bcat2 KO primary adipocytes.

(a) Bcat2 expression is induced along with the SVF-derived primary adipocytes. Bcat2 protein levels as indicated were quantified and normalized against Tubulin. n=1. (b) Bcat1 is not detectable in primary adipocytes. Bcat1 protein levels as indicated were quantified and normalized against Tubulin. n=1. (c) Bcat2 KO increases the mRNA levels of thermogenesis associated genes. n=3. (d) Bcat2 putting back decreases Ucp1 protein expression and mRNA expression of Ucp1, Cidea and Cox8b. Bcat2 was put back by adenovirus carrying Bcat2. Ucp1 protein levels as indicated were quantified and normalized against Tubulin. n=3 independent cultures. (e) Bcat2 KO has no effect on primary adipocytes differentiation rate. Oil-red O staining was performed with mature adipocytes at day 6. Representative result from 2 independent biological experiments. (f) Bcat2 KO has no effect on adipogenesis markers. Fabp4 protein levels as indicated were quantified and normalized against Tubulin. n=3 independent cultures. (g) BCKA downregulates Cidea expression in a dose dependent manner in Bcat2 KO adipocytes. Cidea protein levels as indicated were quantified and normalized against Tubulin. Representative result from 2 independent biological experiments. (h) Bcat2 KO has no effect on the protein levels of Prdm16, Pparγ and Pgc1α. The indicated protein levels were quantified and normalized against Tubulin. n=2 independent cultures. Representative result from 2 independent biological experiments. (i) KO Bcat2 enhances the binding between Pparγ and Prdm16. Pparγ pulled down by Prdm16 was normalized against immunoprecipitated Prdm16. Representative result from 2 independent biological experiments. (j-k). Long term treatment of BCKA or Ac-CoA inhibited Ucp1 expression level. SVF cells were induced to adipocytes with BCKA, Acetyl coenzyme A sodium salt 2-Methylbutyryl-L-carnitine (L-carnitine), Isovaleryl-L-carnitine (IL-carnitine), Isobutyryl-L-carnitine (V-carnitine). +: 0.2 mM, ++: 0.4 mM. Of note, adipocytes were treated with indicated metabolites from induction on day 0 to mature on day 7. Ucp1 protein levels as indicated were quantified and normalized against Tubulin. Representative result from 2 biological experiments. Bars and error bars represent mean values and SDs, with biologically individual data points shown. P values are derived from unpaired, two-tailed t-test.

Source data

Extended Data Fig. 5 Acetylation of Prdm16 in Bcat2 KO primary adipocytes.

(a) Acetate at high but not low doses reverse the effects of Bcat2 KO in mature adipocytes. The acetylation level of Prdm16 as indicated was normalized against immunoprecipitated Prdm16. Representative result from 2 independent biological experiments. (b) TSA increase the acetylation level of Prdm16 but not Pparγ, Pgc1a. TSA: trichostatin A, 8hr with 10uM. NAM: nicotinamide, 8hr with 3mM. N+T: both NAM and TSA. The acetylation level of indicated protein was normalized against immunoprecipitated indicated protein, respectively. (c) Hdac6 decreases the acetylation level of exogeneous Prdm16. Flag-prdm16 and HA-Hdacs were overexpressed in 293T cells. The acetylation level of Prdm16 as indicated was normalized against immunoprecipitated Prdm16-flag. Representative result from 3 independent biological experiments. (d) P300 increases the acetylation level of exogeneous Prdm16. Flag-prdm16 and HATs were overexpressed in 293T cells. The acetylation level of Prdm16 as indicated was normalized against immunoprecipitated Prdm16-flag. Representative result from 3 independent biological experiments. (e) KO Bcat2 decreases the acetylation level of exogeneous Prdm16. The acetylation level of Prdm16 as indicated was normalized against immunoprecipitated Prdm16-flag. Representative result from 2 independent biological experiments. (f) K915 of Prdm16 is conserved in indicated species. (g) K915R mutation abolishes the acetylation level of Prdm16 induced by TSA. The acetylation level of Prdm16 as indicated was normalized against immunoprecipitated Prdm16-flag. Representative result from 2 independent biological experiments. (h) Prdm16K915Q weakens its effect on Ucp1 and Cidea expression. Prdm16WT or Prdm16K915Q mutant was overexpressed into Bcat2 KO adipocytes. Ucp1 protein levels as indicated were quantified and normalized against immunoprecipitated Prdm16WT or Prdm16K915Q protein. n=3 independent cultures. (i) Prdm16K915Q suppresses oxygen consumption rate (OCR). n=3 independent cultures. Bars and error bars represent mean values and SDs, with biologically individual data points shown. P values are derived from unpaired, two-tailed t-test.

Source data

Extended Data Fig. 6 BCKA rescue phenotype induced by Bcat2 KO.

(a) BCKA (KMV and KIC, not including KIV) was supplemented in drinking water at 0.5,1 and 2 mg/ml. Body weight data are showed under HFD. n=5 mice. (b) BCKA supplement doesn’t affect body weight. n=5 mice. (c) BCKA supplement doesn’t affect water intake. Representative mean value from 5 mice in a cage. (d) BCKA supplement doesn’t affect food intake. Representative mean value from 5 mice in a cage. (e) BCKA (KMV and KIC, not including KIV) supplement increase KMV and KIC concentrations in serum. n=3 mice. (f) BCKA supplement rescues body weight. n=5 mice. (g) BCKA restore Bcat2 KO-decreased glucose intolerance and insulin tolerance. n=5 mice. (h) BCKA supplement rescues fatty liver. Representative result from 4 mice. (i) BCKA supplement increase KMV and KIC concentrations in iWAT. n=3 mice. (j) BCKA restore Bcat2 KO-increased Cidea expression in iWAT. Cidea protein levels as indicated were quantified and normalized against Tubulin. n=3 mice. (k) The acetylation level of Prdm16 from BAT. The acetylation level of Prdm16 was normalized against immunoprecipitated Prdm16 protein as indicated. n=3 mice. (l) Ac-CoA level decreases in BAT from Bcat2 Adipose KO mice. n=3 biological independent experiments. (m-n) Ucp1, Prdm16 and Pgc1α expression levels of iWAT and BAT. The protein levels as indicated were quantified and normalized against Tubulin. n=3 mice (m) and n=6 mice (n). Bars and error bars represent mean values and SDs, with biologically individual data points shown. P values are derived from unpaired, two-tailed t-test (except a and g), or repeated-measures 2-way ANOVA followed by Sidak’s multiple-comparison test (Fig. a and g).

Source data

Extended Data Fig. 7 Screening FDA approved drug library for BCAT2 inhibitor.

Sketch map for screening BCAT2 inhibitor from FDA proved library. (b) Repeat data of top 8 compounds from the first-round screen. n=3 biological independent experiments. (c) TEL inhibits human BCAT2 enzyme activity. OD of NADH changes was shown in left panel. BCAT protein was purified from E.Coli and incubated with TEL for 1h, followed by measuring enzyme activity. n=3 biological independent experiments. (d) Other compounds from Sartan catalogue have no effect on BCAT2 activity. n=3 biological independent experiments. (e) TEL inhibits mouse Bcat2 enzyme activity. OD of NADH changes was shown in left panel. NIH3T3 cells transfected with Flag-Bcat2 were treated with TEL for 24h, followed by IP Flag and measuring enzyme activity. n=3 biological independent experiments. Bars and error bars represent mean values and SDs, with biologically individual data points shown. P values are derived from unpaired, two-tailed t-test.

Source data

Extended Data Fig. 8 TEL promotes iWAT browning.

(a) TEL upregulates Cidea expression in primary adipocytes and BCKA rescues this effect. Ucp1 protein levels as indicated were quantified and normalized against Tubulin. n=2 independent cultures. (b) Bcat2 over-expression attenuates the effect of TEL on Ucp1. Ucp1 protein levels as indicated were quantified and normalized against Tubulin. Representative result from 2 independent experiments. (c) TEL reduces acetyl-CoA levels in primary adipocytes. n=3 biological independent experiments. (d) Computed tomography imaging showed TEL prevents mice from HFD induced fate tissue weight gain. n=3 mice. (e) TEL has no effect on food intake. Representative mean value from 5 mice in a cage. (f-g) GTT and ITT from HFD and telmisartan group mice. n=6 mice. (h) Serum markers as indicated from HFD and telmisartan group mice. n=7 mice. (i-j) TEL protects mice from HFD induced hypertension and tachycardia. n=6 mice. (k) TEL has no effect on other main organ weights. n=7 mice. (l) TEL prevents HFD-induced fatty liver. Representative result from 4 mice. (m) In vivo metabolism data from HFD and TEL group mice. n=5 mice. Bars and error bars represent mean values and SDs, with biologically individual data points shown. P values are derived from unpaired, two-tailed t-test (c, d, h-j, m-n), or repeated-measures 2-way ANOVA followed by Sidak’s multiple-comparison test (f, g).

Source data

Extended Data Fig. 9 TEL promotes iWAT browning and energy expenditure.

(a) Energy expenditure was analyzed by ANCOVA, with body weight included as a covariate. n=5 mice. (b-c) TEL increases body temperature under cold exposure but not room temperature. The representative rectal temperatures were shown. n=5 (Control) and n=6 (TEL). (d) Temperature at interscapular area of TEL treated mice is significantly higher than that of control mice showed by thermal infrared imaging. n=4 mice. (e) TEL upregulates thermogenesis related gene expression. n=6. (f) TEL increases Cidea expression in iWAT. Cidea protein levels as indicated were quantified and normalized against Tubulin. n=3. (g) TEL increases Ucp1 expression in BAT. Representative images of Ucp1 immunohistochemistry (IHC) in BAT from mice fed with HFD. Representative result from 4 mice. (h) TEL decreases body weight of obesity mice. n=5 mice. (i) GTT and ITT data from HFD and telmisartan group. n=5 mice. (j) Fat tissue weights of HFD and TEL group mice. n=6 mice. (k) TEL upregulates Ucp1 expression. TEL was given post obesity. Ucp1 protein levels as indicated were quantified and normalized against Tubulin. n=5. Bars and error bars represent mean values and SDs, with biologically individual data points shown. P values are derived from unpaired, two-tailed t-test (b, d, e, j), or repeated-measures 2-way ANOVA followed by Sidak’s multiple-comparison test (c, h, i). Energy expenditure in Fig.a was analyzed by ANCOVA, with body weight included as a covariate.

Source data

Extended Data Fig. 10 BCKA reverses the lean phenotype conferred by TEL.

(a)Body weight gain from mice as indicated groups. n=5 mice. (b) GTT and ITT data from mice as indicated groups. n=5 mice. (c) HE staining of liver histology from mice as indicated groups. Representative result from 4 mice. (d) Ucp1 and Cidea expression for iWAT from mice as indicated groups. Ucp1 and Cidea protein levels as indicated were quantified and normalized against Tubulin. n=3 mice. Bars and error bars represent mean values and SDs, with biologically individual data points shown. n, cohort size. P values are derived from unpaired, two-tailed t-test (a), or repeated-measures 2-way ANOVA followed by Sidak’s multiple-comparison test (b).

Source data

Supplementary information

Supplementary Information

Supplementary Table 1, primers used in this study.

Reporting Summary

Source data

Source Data Fig. 1

Statistical source data.

Source Data Fig. 2

Statistical source data.

Source Data Fig. 2

Unprocessed western blots.

Source Data Fig. 3

Statistical source data.

Source Data Fig. 3

Unprocessed western blots.

Source Data Fig. 4

Statistical source data.

Source Data Fig. 4

Unprocessed Western Blots.

Source Data Fig. 5

Statistical Source Data

Source Data Fig. 6

Statistical Source Data.

Source Data Fig. 6

Unprocessed western blots.

Source Data Fig. 7

Statistical source data.

Source Data Extended Data Fig. 1

Statistical source data.

Source Data Extended Data Fig. 1

Unprocessed western blots.

Source Data Extended Data Fig. 2

Statistical source data.

Source Data Extended Data Fig. 2

Unprocessed western blots.

Source Data Extended Data Fig. 3

Statistical source data.

Source Data Extended Data Fig. 3

Unprocessed western blots.

Source Data Extended Data Fig. 4

Statistical source data.

Source Data Extended Data Fig. 4

Source Data Extended Data Fig. 5

Statistical source data.

Source Data Extended Data Fig. 5

Unprocessed western blots.

Source Data Extended Data Fig. 6

Statistical source data.

Source Data Extended Data Fig. 6

Unprocessed western blots.

Source Data Extended Data Fig. 7

Statistical source data.

Source Data Extended Data Fig. 8

Statistical source data.

Source Data Extended Data Fig. 8

Unprocessed western blots.

Source Data Extended Data Fig. 9

Statistical Source Data.

Source Data Extended Data Fig. 9

Unprocessed Western Blots.

Source Data Extended Data Fig. 10

Statistical source data.

Source Data Extended Data Fig. 10

Unprocessed western blots.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ma, QX., Zhu, WY., Lu, XC. et al. BCAA–BCKA axis regulates WAT browning through acetylation of PRDM16. Nat Metab 4, 106–122 (2022). https://doi.org/10.1038/s42255-021-00520-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s42255-021-00520-6

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing