Calpains Orchestrate Secretion of Annexin-containing Microvesicles During Membrane Repair

Overview

Journal bioRxiv

Date 2024 Sep 16

PMID 39282443

Authors

Justin Krish Williams

Jordan Matthew Ngo

Abinayaa Murugupandiyan

Dorothy E Croall

H Criss Hartzell

Randy Schekman

Affiliations

Soon will be listed here.

Abstract

Microvesicles (MVs) are membrane-enclosed, plasma membrane-derived particles released by cells from all branches of life. MVs have utility as disease biomarkers and may participate in intercellular communication; however, physiological processes that induce their secretion are not known. Here, we isolate and characterize annexin-containing MVs and show that these vesicles are secreted in response to the calcium influx caused by membrane damage. The annexins in these vesicles are cleaved by calpains. After plasma membrane injury, cytoplasmic calcium-bound annexins are rapidly recruited to the plasma membrane and form a scab-like structure at the lesion. In a second phase, recruited annexins are cleaved by calpains-1/2, disabling membrane scabbing. Cleavage promotes annexin secretion within MVs. Our data supports a new model of plasma membrane repair, where calpains relax annexin-membrane aggregates in the lesion repair scab, allowing secretion of damaged membrane and annexins as MVs. We anticipate that cells experiencing plasma membrane damage, including muscle and metastatic cancer cells, secrete these MVs at elevated levels.

References

Muenchbach M, DellAmbrogio M, Gazzotti P . Proteolysis of liver plectin by mu-calpain. Biochem Biophys Res Commun. 1998; 249(2):304-6. DOI: 10.1006/bbrc.1998.9138. View

Fox J, Austin C, Reynolds C, Steffen P . Evidence that agonist-induced activation of calpain causes the shedding of procoagulant-containing microvesicles from the membrane of aggregating platelets. J Biol Chem. 1991; 266(20):13289-95. View

Jimenez A, Maiuri P, Lafaurie-Janvore J, Divoux S, Piel M, Perez F . ESCRT machinery is required for plasma membrane repair. Science. 2014; 343(6174):1247136. DOI: 10.1126/science.1247136. View

Lin Y, Chipot C, Scheuring S . Annexin-V stabilizes membrane defects by inducing lipid phase transition. Nat Commun. 2020; 11(1):230. PMC: 6957514. DOI: 10.1038/s41467-019-14045-w. View

Golia M, Gabrielli M, Verderio C . P2X Receptor and Extracellular Vesicle Release. Int J Mol Sci. 2023; 24(12). PMC: 10298639. DOI: 10.3390/ijms24129805. View

Sonder S, Boye T, Tolle R, Dengjel J, Maeda K, Jaattela M . Annexin A7 is required for ESCRT III-mediated plasma membrane repair. Sci Rep. 2019; 9(1):6726. PMC: 6491720. DOI: 10.1038/s41598-019-43143-4. View

Kearney K, Ariens R, Macrae F . The Role of Fibrin(ogen) in Wound Healing and Infection Control. Semin Thromb Hemost. 2021; 48(2):174-187. DOI: 10.1055/s-0041-1732467. View

Shapovalov I, Harper D, Greer P . Calpain as a therapeutic target in cancer. Expert Opin Ther Targets. 2022; 26(3):217-231. DOI: 10.1080/14728222.2022.2047178. View

Bharadwaj A, Bydoun M, Holloway R, Waisman D . Annexin A2 heterotetramer: structure and function. Int J Mol Sci. 2013; 14(3):6259-305. PMC: 3634455. DOI: 10.3390/ijms14036259. View

10.

Hanna R, Campbell R, Davies P . Calcium-bound structure of calpain and its mechanism of inhibition by calpastatin. Nature. 2008; 456(7220):409-12. DOI: 10.1038/nature07451. View

11.

Krish Williams J, Ngo J, Lehman I, Schekman R . Annexin A6 mediates calcium-dependent exosome secretion during plasma membrane repair. Elife. 2023; 12. PMC: 10241516. DOI: 10.7554/eLife.86556. View

12.

McNeil P, Khakee R . Disruptions of muscle fiber plasma membranes. Role in exercise-induced damage. Am J Pathol. 1992; 140(5):1097-109. PMC: 1886518. View

13.

Leloup L, Shao H, Bae Y, Deasy B, Stolz D, Roy P . m-Calpain activation is regulated by its membrane localization and by its binding to phosphatidylinositol 4,5-bisphosphate. J Biol Chem. 2010; 285(43):33549-33566. PMC: 2963356. DOI: 10.1074/jbc.M110.123604. View

14.

Corrotte M, Castro-Gomes T, Koushik A, Andrews N . Approaches for plasma membrane wounding and assessment of lysosome-mediated repair responses. Methods Cell Biol. 2015; 126:139-58. PMC: 4739812. DOI: 10.1016/bs.mcb.2014.11.009. View

15.

Mellgren R, Miyake K, Kramerova I, Spencer M, Bourg N, Bartoli M . Calcium-dependent plasma membrane repair requires m- or mu-calpain, but not calpain-3, the proteasome, or caspases. Biochim Biophys Acta. 2009; 1793(12):1886-93. PMC: 2787696. DOI: 10.1016/j.bbamcr.2009.09.013. View

16.

Romero M, Keyel M, Shi G, Bhattacharjee P, Roth R, Heuser J . Intrinsic repair protects cells from pore-forming toxins by microvesicle shedding. Cell Death Differ. 2017; 24(5):798-808. PMC: 5423106. DOI: 10.1038/cdd.2017.11. View

17.

Williams S, Milne I, Bagley C, Gamble J, Vadas M, Pitson S . A proinflammatory role for proteolytically cleaved annexin A1 in neutrophil transendothelial migration. J Immunol. 2010; 185(5):3057-63. DOI: 10.4049/jimmunol.1000119. View

18.

Rezvanpour A, Santamaria-Kisiel L, Shaw G . The S100A10-annexin A2 complex provides a novel asymmetric platform for membrane repair. J Biol Chem. 2011; 286(46):40174-83. PMC: 3220529. DOI: 10.1074/jbc.M111.244038. View

19.

Reber S, Mechtersheimer J, Nasif S, Benitez J, Colombo M, Domanski M . CRISPR-Trap: a clean approach for the generation of gene knockouts and gene replacements in human cells. Mol Biol Cell. 2017; 29(2):75-83. PMC: 5909934. DOI: 10.1091/mbc.E17-05-0288. View

20.

Prislusky M, Lam J, Contreras V, Ng M, Chamberlain M, Pathak-Sharma S . The septin cytoskeleton is required for plasma membrane repair. EMBO Rep. 2024; 25(9):3870-3895. PMC: 11387490. DOI: 10.1038/s44319-024-00195-6. View