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.

  • Correspondence
  • Published:

PKM2 controls the degranulation of secondary and tertiary granules in neutrophils by phosphorylating SNAP-23

Cellular & Molecular Immunology volume 18, pages 2048–2050 (2021)Cite this article

Subjects

Polymorphonuclear neutrophils (PMNs) possess multiple distinct types of cytoplasmic granules: secondary granules, tertiary granules, and azurophil granules [1]. It has been suggested that PMN degranulation is controlled by exocytotic machinery comprising membrane-specific SNARE proteins, soluble N-ethylmaleimide sensitive factor (NSF) and soluble NSF-attaching proteins [2]. Indeed, different SNARE proteins, SNAP-23 and SNAP-25, have been identified in PMNs and play essential roles in PMN degranulation [3]. However, the mechanisms governing the activation of these SNARE proteins during PMN degranulation remain unknown.

To further test whether PKM2 is involved in selective degranulation in the PMN, we examined whether fMLP-stimulated PMN degranulation is altered by reagents that modulate PKM2 phosphorylation and dimeric or tetrameric formation. In this experiment, PMNs were treated with pTyr, a phosphotyrosine peptide, to promote PKM2 phosphorylation and dimeric formation [5] and DASA-58 or TEPP-46 to enhance PKM2 tetrameric formation. Treatment with DASA-58 or TEPP-46 markedly reduced fMLP-stimulated PMN degranulation of specific and tertiary granules (Fig. 1c and Supplementary Fig. 2). In contrast, the pTyr peptide promoted the degranulation of secondary and tertiary granules in the fMLP-stimulated PMNs (Fig. 1c and Supplementary Fig. 2). In fact, pTyr treatment enhanced PMN degranulation of secondary and tertiary granules in the absence of fMLP stimulation. These results suggest that phosphorylated PKM2 promotes degranulation of PMN secondary and tertiary granules.

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

References

  1. Borregaard N, Sorensen OE, Theilgaard-Monch K. Neutrophil granules: a library of innate immunity proteins. Trends Immunol. 2007;28:340–5.

    Article  CAS  Google Scholar 

  2. Rothman JE. Mechanisms of intracellular protein transport. Nature. 1994;372:55–63.

    Article  CAS  Google Scholar 

  3. Martin-Martin B, Nabokina SM, Blasi J, Lazo PA, Mollinedo F. Involvement of SNAP-23 and syntaxin 6 in human neutrophil exocytosis. Blood. 2000;96:2574–83.

    Article  CAS  Google Scholar 

  4. Rodriguez-Espinosa O, Rojas-Espinosa O, Moreno-Altamirano MM, Lopez-Villegas EO, Sanchez-Garcia FJ. Metabolic requirements for neutrophil extracellular traps formation. Immunology. 2015;145:213–24.

    Article  CAS  Google Scholar 

  5. Christofk HR, Vander Heiden MG, Wu N, Asara JM, Cantley LC. Pyruvate kinase M2 is a phosphotyrosine-binding protein. Nature. 2008;452:181–6.

    Article  CAS  Google Scholar 

  6. Kallquist L, Hansson M, Persson AM, Janssen H, Calafat J, Tapper H, et al. The tetraspanin CD63 is involved in granule targeting of neutrophil elastase. Blood. 2008;112:3444–54.

    Article  Google Scholar 

  7. Wei Y, Wang D, Jin F, Bian Z, Li L, Liang H, et al. Pyruvate kinase type M2 promotes tumour cell exosome release via phosphorylating synaptosome-associated protein 23. Nat. Commun. 2017;8:14041.

    Article  CAS  Google Scholar 

  8. Yang W, Xia Y, Hawke D, Li X, Liang J, Xing D, et al. PKM2 phosphorylates histone H3 and promotes gene transcription and tumorigenesis. Cell. 2012;150:685–96.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from the Ministry of Science and Technology of China (2018YFA0507100), the National Natural Science Foundation of China (31670917, 31770981, 32000634, and 81803626), the Natural Science Foundation of Jiangsu Province (BK20180556), and the fellowship of China Postdoctoral Science Foundation (2018M630642).

Author information

Author notes

  1. These authors contributed equally: Xinlei Sun, Dong Wang.

Authors and Affiliations

  1. School of Life Science and Technology, China Pharmaceutical University, Nanjing, Jiangsu, China

    Xinlei Sun, Shuang Qu, Hongwei Liang & Ke Zen

  2. Department of Pharmacology, School of Pharmacy, China Pharmaceutical University, Nanjing, Jiangsu, China

    Dong Wang

  3. School of Life Sciences, Nanjing University, Nanjing, Jiangsu, China

    Yan Wang, Fujie Shi, Shuang Qu & Ke Zen

  4. Department of Gastroenterology and Hepatology, Jinling Hospital, Medical School of Nanjing University, Nanjing, China

    Lei Ye

Authors
  1. Xinlei Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Dong Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yan Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Lei Ye
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Fujie Shi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Shuang Qu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Hongwei Liang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Ke Zen
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

KZ designed the experiments. X.S., D.W., Y.W., L.Y., Q.S., F.S., and H.L. performed the experiments and analyzed the results. X.S. and K.Z. wrote the paper.

Corresponding author

Correspondence to Ke Zen.

Ethics declarations

Competing interests

The authors declare no competing interests.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sun, X., Wang, D., Wang, Y. et al. PKM2 controls the degranulation of secondary and tertiary granules in neutrophils by phosphorylating SNAP-23. Cell Mol Immunol 18, 2048–2050 (2021). https://doi.org/10.1038/s41423-021-00718-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41423-021-00718-4

Search

Advanced search

Quick links