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Cited by 36Glycosylation and Glycoproteins ResearchLipid Membrane Structure and BehaviorCellular transport and secretion

Human Sialidase Neu3 is S-Acylated and Behaves Like an Integral Membrane Protein

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Rodriguez-Walker, Macarena, Daniotti, Jose L.·2017·Scientific Reports
DOI 10.1038/s41598-017-04488-wPMC PMC5482835
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Abstract

Membrane-bound sialidase Neu3 is involved in the catabolism of glycoconjugates, and plays crucial roles in numerous biological processes. Since the mechanism of its association with membranes is still not completely understood, the aim of this work was to provide further information regarding this aspect. Human Neu3 was found to be associated with the plasma membrane and endomembranes, and it was not released from the lipid bilayer under conditions that typically release peripheral membrane proteins. By different experimental approaches, we demonstrated that its C-terminus is exposed to the cytosol while another portion of the protein is exposed to the extracellular space, suggesting that Neu3 possesses the features of a transmembrane protein. However, in silico analysis and homology modeling predicted that the sialidase does not contain any α-helical transmembrane segment and shares the same β-propeller fold typical of viral and bacterial sialidases. Additionally, we found that Neu3 is S-acylated. Since this post-translational modification is restricted to the cytosolic side of membranes, this finding strongly supports the idea that Neu3 may contain a cytosolic-exposed domain. Although it remains to be determined exactly how this sialidase crosses the lipid bilayer, this study provides new insights about membrane association and topology of Neu3.

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  • The NEU1 oligomerization mode is unrelated to viral sialidase tetramers, disulfide-mediated NEU3 dimers, or the various lattice interfaces observed in NEU2 crystal structures.

    “The NEU1 oligomerization mode is unrelated to viral sialidase tetramers (62), disulfide-mediated NEU3 dimers (9), or the various lattice interfaces observed in NEU2 crystal structures (11, 75, 76), raising the possibility that NEU1 self-association may be required for complex formation with CTSA.”

    From Structure of the immunoregulatory sialidase NEU1 (2023) · cites this paper as [9]

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  1. [1]Miyagi T Yamaguchi K Mammalian sialidases: physiological and pathological roles in cellular functions Glycobiology 2012 22 880 896 10.1093/glycob/cws057 22377912 · DOI
  2. [2]Monti E Sialidases in vertebrates: a family of enzymes tailored for several cell functions Adv Carbohydr Chem Biochem 2010 64 403 479 10.1016/S0065-2318(10)64007-3 20837202 · DOI
  3. [3]Miyagi T Molecular cloning and characterization of a plasma membrane-associated sialidase specific for gangliosides J Biol Chem 1999 274 5004 5011 10.1074/jbc.274.8.5004 9988745 · DOI
  4. [4]Anastasia L NEU3 sialidase strictly modulates GM3 levels in skeletal myoblasts C2C12 thus favoring their differentiation and protecting them from apoptosis J Biol Chem 2008 283 36265 36271 10.1074/jbc.M805755200 18945680 PMC2662296 · DOI
  5. [5]Jia F Howlader MA Cairo CW Integrin-mediated cell migration is blocked by inhibitors of human neuraminidase Biochim Biophys Acta 2016 1861 1170 1179 10.1016/j.bbalip.2016.06.013 27344026 · DOI
  6. [6]Papini N MmNEU3 sialidase over-expression in C2C12 myoblasts delays differentiation and induces hypertrophic myotube formation J Cell Biochem 2012 113 2967 2978 10.1002/jcb.24174 22552967 · DOI
  7. [7]Wada T A crucial role of plasma membrane-associated sialidase in the survival of human cancer cells Oncogene 2007 26 2483 2490 10.1038/sj.onc.1210341 17334392 · DOI
  8. [8]Miyagi T Wada T Yamaguchi K Roles of plasma membrane-associated sialidase NEU3 in human cancers Biochim Biophys Acta 2008 1780 532 537 10.1016/j.bbagen.2007.09.016 18023981 · DOI
  9. [9]Rodriguez-Walker M Vilcaes AA Garbarino-Pico E Daniotti JL Role of plasma-membrane-bound sialidase NEU3 in clathrin-mediated endocytosis Biochem J 2015 470 131 144 10.1042/BJ20141550 26251452 · DOI
  10. [10]Yamamoto K Potentiation of epidermal growth factor-mediated oncogenic transformation by sialidase NEU3 leading to Src activation PLoS One 2015 10 e0120578 10.1371/journal.pone.0120578 25803810 PMC4372364 · DOI
  11. [11]Wang Y A close association of the ganglioside-specific sialidase Neu3 with caveolin in membrane microdomains J Biol Chem 2002 277 26252 26259 10.1074/jbc.M110515200 12011038 · DOI
  12. [12]Sasaki A Overexpression of plasma membrane-associated sialidase attenuates insulin signaling in transgenic mice J Biol Chem 2003 278 27896 27902 10.1074/jbc.M212200200 12730204 · DOI
  13. [13]Kato K Plasma-membrane-associated sialidase (NEU3) differentially regulates integrin-mediated cell proliferation through laminin- and fibronectin-derived signalling Biochem J 2006 394 647 656 10.1042/BJ20050737 16241905 PMC1383714 · DOI
  14. [14]Smutova V Structural basis for substrate specificity of mammalian neuraminidases PLoS One 2014 9 e106320 10.1371/journal.pone.0106320 25222608 PMC4164519 · DOI
  15. [15]Mozzi A NEU3 activity enhances EGFR activation without affecting EGFR expression and acts on its sialylation levels Glycobiology 2015 25 855 868 10.1093/glycob/cwv026 25922362 · DOI
  16. [16]Zanchetti G Sialidase NEU3 is a peripheral membrane protein localized on the cell surface and in endosomal structures Biochem J 2007 408 211 219 10.1042/BJ20070503 17708748 PMC2267352 · DOI
  17. [17]Cirillo F NEU3 Sialidase Protein Interactors in the Plasma Membrane and in the Endosomes J Biol Chem 2016 291 10615 10624 10.1074/jbc.M116.719518 26987901 PMC4865910 · DOI
  18. [18]Chavas LM Crystal structure of the human cytosolic sialidase Neu2. Evidence for the dynamic nature of substrate recognition J Biol Chem 2005 280 469 475 10.1074/jbc.M411506200 15501818 · DOI · Paper page
  19. [19]Varghese JN Laver WG Colman PM Structure of the influenza virus glycoprotein antigen neuraminidase at 2.9 A resolution Nature 1983 303 35 40 10.1038/303035a0 6843658 · DOI · Paper page
  20. [20]Crennell SJ Garman EF Laver WG Vimr ER Taylor GL Crystal structure of a bacterial sialidase (from Salmonella typhimurium LT2) shows the same fold as an influenza virus neuraminidase Proc Natl Acad Sci USA 1993 90 9852 9856 10.1073/pnas.90.21.9852 8234325 PMC47670 · DOI · Paper page
  21. [21]Magesh S Suzuki T Miyagi T Ishida H Kiso M Homology modeling of human sialidase enzymes NEU1, NEU3 and NEU4 based on the crystal structure of NEU2: hints for the design of selective NEU3 inhibitors J Mol Graph Model 2006 25 196 207 10.1016/j.jmgm.2005.12.006 16427342 · DOI
  22. [22]Crespo PM Zurita AR Daniotti JL Effect of gangliosides on the distribution of a glycosylphosphatidylinositol-anchored protein in plasma membrane from Chinese hamster ovary-K1 cells J Biol Chem 2002 277 44731 44739 10.1074/jbc.M204604200 12237294 · DOI
  23. [23]Papini N The plasma membrane-associated sialidase MmNEU3 modifies the ganglioside pattern of adjacent cells supporting its involvement in cell-to-cell interactions J Biol Chem 2004 279 16989 16995 10.1074/jbc.M400881200 14970224 · DOI
  24. [24]Gomez GA Daniotti JL Electrical properties of plasma membrane modulate subcellular distribution of K-Ras FEBS J 2007 274 2210 2228 10.1111/j.1742-4658.2007.05758.x 17388810 · DOI
  25. [25]Root KT Plucinsky SM Glover KJ Recent progress in the topology, structure, and oligomerization of caveolin: a building block of caveolae Curr Top Membr 2015 75 305 336 10.1016/bs.ctm.2015.03.007 26015287 · DOI
  26. [26]Kyte J Doolittle RF A simple method for displaying the hydropathic character of a protein J Mol Biol 1982 157 105 132 10.1016/0022-2836(82)90515-0 7108955 · DOI
  27. [27]Monti E Preti A Venerando B Borsani G Recent development in mammalian sialidase molecular biology Neurochem Res 2002 27 649 663 10.1023/A:1020276000901 12374200 · DOI
  28. [28]Chamberlain LH Shipston MJ The physiology of protein S-acylation Physiol Rev 2015 95 341 376 10.1152/physrev.00032.2014 25834228 PMC4551212 · DOI
  29. [29]Dietzen DJ Hastings WR Lublin DM Caveolin is palmitoylated on multiple cysteine residues. Palmitoylation is not necessary for localization of caveolin to caveolae J Biol Chem 1995 270 6838 6842 10.1074/jbc.270.12.6838 7896831 · DOI
  30. [30]Nadolski MJ Linder ME Protein lipidation FEBS J 2007 274 5202 5210 10.1111/j.1742-4658.2007.06056.x 17892486 · DOI
  31. [31]Trenchi A Gomez GA Daniotti JL Dual acylation is required for trafficking of growth-associated protein-43 (GAP-43) to endosomal recycling compartment via an Arf6-associated endocytic vesicular pathway Biochem J 2009 421 357 369 10.1042/BJ20090484 19442238 · DOI
  32. [32]Albohy A Li MD Zheng RB Zou C Cairo CW Insight into substrate recognition and catalysis by the human neuraminidase 3 (NEU3) through molecular modeling and site-directed mutagenesis Glycobiology 2010 20 1127 1138 10.1093/glycob/cwq077 20511247 · DOI
  33. [33]Wang Y Yamaguchi K Shimada Y Zhao X Miyagi T Site-directed mutagenesis of human membrane-associated ganglioside sialidase: identification of amino-acid residues contributing to substrate specificity Eur J Biochem 2001 268 2201 2208 10.1046/j.1432-1327.2001.02069.x 11298736 · DOI
  34. [34]Roggentin P Schauer R Hoyer LL Vimr ER The sialidase superfamily and its spread by horizontal gene transfer Mol Microbiol 1993 9 915 921 10.1111/j.1365-2958.1993.tb01221.x 7934919 · DOI
  35. [35]Albohy A Mohan S Zheng RB Pinto BM Cairo CW Inhibitor selectivity of a new class of oseltamivir analogs against viral neuraminidase over human neuraminidase enzymes Bioorg Med Chem 2011 19 2817 2822 10.1016/j.bmc.2011.03.039 21489803 · DOI
  36. [36]Magesh S Design, synthesis, and biological evaluation of human sialidase inhibitors. Part 1: selective inhibitors of lysosomal sialidase (NEU1) Bioorg Med Chem Lett 2008 18 532 537 10.1016/j.bmcl.2007.11.084 18068975 · DOI
  37. [37]Zhang Y Identification of selective inhibitors for human neuraminidase isoenzymes using C4, C7-modified 2-deoxy-2,3-didehydro-N-acetylneuraminic acid (DANA) analogues J Med Chem 2013 56 2948 2958 10.1021/jm301892f 23530623 · DOI
  38. [38]Otto GP Nichols BJ The roles of flotillin microdomains–endocytosis and beyond J Cell Sci 2011 124 3933 3940 10.1242/jcs.092015 22194304 · DOI
  39. [39]Lundmark R Carlsson SR Sorting nexin 9 participates in clathrin-mediated endocytosis through interactions with the core components J Biol Chem 2003 278 46772 46781 10.1074/jbc.M307334200 12952949 · DOI
  40. [40]Sato K Nakano A Mechanisms of COPII vesicle formation and protein sorting FEBS Lett 2007 581 2076 2082 10.1016/j.febslet.2007.01.091 17316621 · DOI
  41. [41]Shiozaki K Phosphatidic acid-mediated activation and translocation to the cell surface of sialidase NEU3, promoting signaling for cell migration FASEB J 2015 29 2099 2111 10.1096/fj.14-262543 25678627 · DOI
  42. [42]Maurice P New Insights into Molecular Organization of Human Neuraminidase-1: Transmembrane Topology and Dimerization Ability Sci Rep 2016 6 38363 10.1038/srep38363 27917893 PMC5137157 · DOI
  43. [43]Petersen TN Brunak S von Heijne G Nielsen H SignalP 4.0: discriminating signal peptides from transmembrane regions Nat Methods 2011 8 785 786 10.1038/nmeth.1701 21959131 · DOI · Paper page
  44. [44]Ruggiero FM Vilcaes AA Iglesias-Bartolomé R Daniotti JL Critical role of evolutionarily conserved glycosylation at Asn211 in the intracellular trafficking and activity of sialyltransferase ST3Gal-II Biochem J 2015 469 83 95 10.1042/BJ20150072 25916169 · DOI
  45. [45]Gomez GA Daniotti JL H-Ras dynamically interacts with recycling endosomes in CHO-K1 cells: involvement of Rab5 and Rab11 in the trafficking of H-Ras to this pericentriolar endocytic compartment J Biol Chem 2005 280 34997 35010 10.1074/jbc.M506256200 16079139 · DOI
  46. [46]Wan J Roth AF Bailey AO Davis NG Palmitoylated proteins: purification and identification Nat Protoc 2007 2 1573 1584 10.1038/nprot.2007.225 17585299 · DOI
  47. [47]Buchan DW Minneci F Nugent TC Bryson K Jones DT Scalable web services for the PSIPRED Protein Analysis Workbench Nucleic Acids Res 2013 41 W349 357 10.1093/nar/gkt381 23748958 PMC3692098 · DOI
  48. [48]Pollastri G McLysaght A Porter: a new, accurate server for protein secondary structure prediction Bioinformatics 2005 21 1719 1720 10.1093/bioinformatics/bti203 15585524 · DOI
  49. [49]Lin K Simossis VA Taylor WR Heringa J A simple and fast secondary structure prediction method using hidden neural networks Bioinformatics 2005 21 152 159 10.1093/bioinformatics/bth487 15377504 · DOI
  50. [50]Drozdetskiy A Cole C Procter J Barton GJ JPred4: a protein secondary structure prediction server Nucleic Acids Res 2015 43 W389 394 10.1093/nar/gkv332 25883141 PMC4489285 · DOI
  51. [51]Schwede T Kopp J Guex N Peitsch MC SWISS-MODEL: An automated protein homology-modeling server Nucleic Acids Res 2003 31 3381 3385 10.1093/nar/gkg520 12824332 PMC168927 · DOI

Cited by

Sorted by citations

Authors
Rodriguez-Walker, M., & Daniotti, J. L.
Affiliations
Consejo Nacional de Investigaciones Científicas y Técnicas (AR)
Corresponding author
Daniotti, Jose L.
ORCID
Daniotti, Jose L.
Venue
Scientific Reports
Publisher
Springer Science and Business Media LLC
Published
2017-06-23
Volume
7
Issue
1
Pages
4167-4167
ISSN
2045-2322 (ISSN-L)
DOI
10.1038/s41598-017-04488-w
PMC
PMC5482835
OpenAlex
W2714698056
Cited by
36
References
0
Citations by year
2026: 2 · 2025: 1 · 2024: 6 · 2023: 4 · 2022: 7 · 2021: 1 · 2020: 7 · 2019: 2 · 2018: 4 · 2017: 2
FWCI
1.74
Type
journal article
Language
en
Open access
Licence
Locations
  • Scientific Reports (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Americanae (AECID Library) (submittedVersion, cc-by-nc-sa)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • LA Referencia (Red Federada de Repositorios Institucionales de Publicaciones Científicas) (submittedVersion, cc-by-nc-sa)
  • Greater South Information System
Funders
  • Mizutani Foundation for Glycoscience (160059)
  • Universidad Nacional de Córdoba (104/15, 203/14)
  • Consejo Nacional de Investigaciones Científicas y Técnicas (PIP 112, PIP 112-20110100930)
  • Agencia Nacional de Promoción Científica y Tecnológica (PICT-2013-0456, 203/14, CONICET)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Glycosylation and Glycoproteins Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Lipid Membrane Structure and Behavior
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
Keywords
S-acylation, membrane association, transmembrane protein topology, plasma membrane, endomembranes, post-translational modification, protein localization
MeSH terms
Acylation, Amino Acid Sequence, Animals, Cricetulus, Disulfides, Cricetinae, Humans, Membrane Proteins, Neuraminidase, CHO Cells, Protein Structure, Secondary, Structural Homology, Protein

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