Page 251 - Haematologica April 2020

The role of neuraminidases in platelet function
This study shows for the first time that NEU1 co- localises with some but not all mitochondria within platelets, while it does not co-localise with LAMP as pre- viously demonstrated.56 In line with our NEU1 findings, NEU4 is located within mitochondria in other nucleated cells16 and NEU4 also translocates to the platelet mem- brane following VWF addition under shear. Moreover, in our hands, NEU1 did not co-localise with β-galactosidase in unstimulated healthy or stimulated platelets, despite previous findings indicating these are both present in lysosomes, and not in platelet α and δ-granules.36 This is in contrast with stored platelets, wherein some diffuse co-localisation of NEU1, with β-galactosidase was observed,22 although this localisation was partial and mostly cytoplasmic. NEU2 however, was co-localised with the α-granule protein P-selectin, which is not sur- prising as P-selectin is sialylated57 and DANA showed a trend towards inhibition of P-selectin expression post- ristocetin treatment.
The findings presented here demonstrate novel roles for
NEU1 and NEU2 in healthy platelets, which are well reg- ulated down-stream of GPIbα following VWF-binding, negatively by calcium (NEU2 only) and increased in pres- ence of fibrinogen. Fibrinogen-binding is required for NEU1 and NEU2 membrane association, enhancing their activity. However when platelet αIIbβ3 is fully occupied with fibrinogen, NEU activity is inhibited.
Acknowledgements
We would like to thank Mikki Diep, Fiona Gardner, Jeannene Moore and Jenny Fisher for assistance with phlebotomy as well as all volunteers for donating blood. We would like to thank Dr. Anja Gerrits for fruitful discussions.
Funding
Australian governments fund the Australian Red Cross Blood Service to provide blood, blood products and services to the Australian community. The Australian & New Zealand society for Blood Transfusion (ANZSBT) provided funding for part of this project.
References
1. Jamieson GA, Okumura T, Hasitz M. Structure and function of platelet glyco- calicin. Thromb haemost. 1980;42(5):1673- 1678.
2. Tsuji T, Tsunehisa S, Watanabe Y, Yamamoto K, Tohyama H, Osawa T. The carbohydrate moiety of human platelet gly- cocalicin. J Biol Chem. 1983;258(10):6335- 6339.
3. Korrel SA, Clemetson KJ, van HH, Kamerling JP, Sixma JJ, Vliegenthart JF. Identification of a tetrasialylated monofuco- sylated tetraantennary N-linked carbohy- drate chain in human platelet glycocalicin. FEBS Lett. 1988;228(2):321-326.
4. King SL, Joshi HJ, Schjoldager KT, et al. Characterizing the O-glycosylation land- scape of human plasma, platelets, and endothelial cells. Blood Aadv. 2017;1(7):429- 442.
5. Grozovsky R, Hoffmeister KM, Falet H. Novel clearance mechanisms of platelets. Curr Opin Hematol. 2010;17(6):585-589.
6. Hoffmeister KM, Felbinger TW, Falet H, et al. The clearance mechanism of chilled blood platelets. Cell. 2003;112(1):87-97.
7. van der Wal DE, Verhoef S, Schutgens RE, Peters M, Wu Y, Akkerman JW. Role of gly- coprotein Ibalpha mobility in platelet func- tion. Thromb Haemost. 2010;103(5):1033- 1043.
8. van der Wal DE, Du VX, Lo KS, Rasmussen JT, Verhoef S, Akkerman JW. Platelet apop- tosis by cold-induced glycoprotein Ibalpha clustering. J Thromb Haemost. 2010;8(11): 2554-2562.
9. Chen W, Druzak SA, Wang Y, et al. Refrigeration-induced binding of von Willebrand factor facilitates fast clearance of refrigerated platelets. Arterioscl Thromb Vasc Biol. 2017;37(12):2271-2279.
10. LiJ,vanderWalDE,ZhuG,etal. Desialylation is a mechanism of Fc-indepen- dent platelet clearance and a therapeutic tar- get in immune thrombocytopenia. Nat Comm. 2015;6:7737.
11. Urbanus RT, van der Wal DE, Koekman CA, et al. Patient autoantibodies induce platelet destruction signals via raft-associated glyco-
protein Ibα and Fc RIIa in immune thrombo- cytopenia. Haematologica. 2013;98(7):e70- e72.
12. Monti E, Bonten E, D'Azzo A, et al. Sialidases in vertebrates: a family of enzymes tailored for several cell functions. Adv Carbohydr Chem Biochem. 2010;64: 403-479.
13. Bonten E, van der Spoel A, Fornerod M, Grosveld G, d'Azzo A. Characterization of human lysosomal neuraminidase defines the molecular basis of the metabolic storage disorder sialidosis. Genes Devel. 1996;10 (24):3156-3169.
14. Mozzi A, Mazzacuva P, Zampella G, Forcella ME, Fusi PA, Monti E. Molecular insight into substrate recognition by human cytosolic sialidase NEU2. Proteins. 2012;80(4):1123-1132.
15. Ha KT, Lee YC, Cho SH, Kim JK, Kim CH. Molecular characterization of membrane type and ganglioside-specific sialidase (Neu3) expressed in E. coli. Molec Cells. 2004;17(2):267-273.
16. Seyrantepe V, Landry K, Trudel S, Hassan JA, Morales CR, Pshezhetsky AV. Neu4, a novel human lysosomal lumen sialidase, confers normal phenotype to sialidosis and galactosialidosis cells. J Biol Chem. 2004;279(35):37021-37029.
17. Bosmann HB, Myers MW, Dehond D, Ball R, Case KR. Mitochondrial autonomy. Sialic acid residues on the surface of isolated rat cerebral cortex and liver mitochondria. J Cell Biol. 1972;55(1):147-160.
18. Abdulkhalek S, Amith SR, Franchuk SL, et al. Neu1 sialidase and matrix metallopro- teinase-9 cross-talk is essential for TOLL- like receptor activation and cellular signal- ing. J Biol Chem. 2011;286(42):36532-36549.
19. Yogalingam G, Bonten EJ, van de Vlekkert D, et al. Neuraminidase 1 is a negative regu- lator of lysosomal exocytosis. Devel Cell. 2008;15(1):74-86.
20. Uemura T, Shiozaki K, Yamaguchi K, et al. Contribution of sialidase NEU1 to suppres- sion of metastasis of human colon cancer cells through desialylation of integrin beta4. Oncogene. 2009;28(9):1218-1229.
21. Kato K, Shiga K, Yamaguchi K, et al. Plasma- membrane-associated sialidase (NEU3) dif- ferentially regulates integrin-mediated cell
proliferation through laminin- and fibronectin-derived signalling. Biochem J. 2006;394(Pt 3):647-656.
22. Jansen AJ, Josefsson EC, Rumjantseva V, et al. Desialylation accelerates platelet clear- ance after refrigeration and initiates GPIbalpha metalloproteinase-mediated cleavage in mice. Blood. 2012;119(5):1263- 1273.
23. Wu KK, Ku CS. Effect of platelet activation on the platelet surface sialic acid. Thromb Res. 1979;14(4-5):697-704.
24. Gitz E, Koekman CA, van den Heuvel DJ, et al. Improved platelet survival after cold stor- age by prevention of Glycoprotein Ibalpha clustering in lipid rafts. Haematologica. 2012;97(12):1873-1881.
25. Bosmann HB. Platelet adhesiveness and aggregation. II. Surface sialic acid, glycopro- tein: N-acetylneuraminic acid transferase, and neuraminidase of human blood platelets. Biochim Biophys Acta. 1972;279 (3):456-474.
26. Potier M, Mameli L, Belisle M, Dallaire L, Melancon SB. Fluorometric assay of neu- raminidase with a sodium (4-methylumbel- liferyl-alpha-D-N-acetylneuraminate) sub- strate. Anal Biochem. 1979;94(2):287-296.
27. Deng W, Xu Y, Chen W, et al. Platelet clear- ance via shear-induced unfolding of a mem- brane mechanoreceptor. Nat Commun. 2016;7:12863.
28. van der Wal DE, Gitz E, Du VX, et al. Arachidonic acid depletion extends survival of cold stored platelets by interfering with [Glycoprotein Ibalpha - 14-3-3zeta] associa- tion. Haematologica. 2012;97(10):1514- 1522.
29. Ravanat C, Strassel C, Hechler B, et al. A central role of GPIb-IX in the procoagulant function of platelets that is independent of the 45-kDa GPIbalpha N-terminal extracel- lular domain. Blood. 2010;116(7):1157-1164.
30. Bergmeier W, Bouvard D, Eble JA, et al. Rhodocytin (aggretin) activates platelets lacking alpha(2)beta(1) integrin, glycopro- tein VI, and the ligand-binding domain of glycoprotein Ibalpha. J Biol Chem. 2001;276(27):25121-25126.
31. Kinlough-Rathbone RL, Perry DW, Rand ML, Packham MA. Responses to aggregat- ing agents after cleavage of GPIb of human
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