Abstract
Sialidases or neuraminidases play important roles in various physiological and pathological processes by cleaving terminal sialic acids (Sias) (desialylation) from the glycans of both glycoproteins and glycolipids. To understand the biological significance of desialylation by sialidases, it is important to investigate enzyme specificity with native substrate in biological membrane of cells. Herein, we report a membrane-mimicking system with liposome ganglioside conjugates containing different lipids for evaluating substrate specificity of sialidase and the lipid effect on the enzyme activity. Briefly, liposomes of phosphatidylcholine (PC) and cholesterol with ganglioside (GM3 or GM1) along with different percentage of phosphatidylserine (PS) or phosphatidylethanolamine (PE) were prepared and characterized. Their desialylation profiles with Arthrobacter ureafaciens (bacterial) sialidase and H1N1 (influenza viral) sialidase were quantified by HPLC method. A diversity of substrate preference was found for both bacterial and viral sialidase to the liposome ganglioside conjugate platform. The apparent Km and Vmax were dependent on the type of lipid. These results indicate that the intrinsic characteristics of the membrane-like system affect the sialidase specificity and activity. This biomimetic substrate provides a better tool for unravelling the substrate specificity and the biological function of sialidases and for screening of functional sialidase inhibitors as well.
Similar content being viewed by others
References
Kelm, S., Schauer, R.: Sialic acids in molecular and cellular interactions. Int. Rev. Cytol.175, 137–240 (1997)
Alley, W.R., Novotny, M.V.: Glycomic analysis of sialic acid linkages in glycans derive from blood serum glycoproteins. J. Proteome Res.9, 3062–3072 (2010)
Schauer, R.: Achievements and challenges of sialic acid research. Glycoconj J.17, 485–499 (2000)
Cohen, M., Varki, A.: The sialome-far more than the sum of its parts. OMICS. 14, 455–464 (2010)
Monti, E., Preti, A., Venerando, B., Borsani, G.: Recent development in mammalian sialidase molecular biology. Neurochem. Res.27, 649–663 (2002)
Hasegawa, T., Sugeno, N., Takeda, A., Kobayashi, M., Kikuchi, A., Furukawa, K., Miyagi, T., Itoyama, Y.: Role of Neu4L sialidase and its substrate ganglioside GD3 in neuronal apoptosis induced by catechol metabolites. FEBS Lett.581, 406–412 (2007)
Sasaki, A., Hata, K., Suzuki, S., Sawada, M., Wada, T., Yamaguchi, K., Obinata, M., Tateno, H., Suzuki, H., Miyagi, T.: Overexpression of plasma membrane-associated sialidase attenuates insulin signaling in transgenic mice. J. Biol. Chem.278, 27896–27902 (2003)
Miyagi, T., Yamaguchi, K.: Mammalian sialidases: physiological and pathological roles in cellular functions. Glycobiology. 22, 880–896 (2012)
Saito, M, Yu, R.K. Biochemistry and Function of Sialidases. In: Rosenberg A. (eds) Biology of the Sialic Acids. 1995.
Lewis, A.L., Lewis, W.G.: Host sialoglycans and bacterial sialidases: a mucosal perspective. Cell Microbiol.14, 1174–1182 (2012)
Szymanski, C.M., Schnaar, R.L., Aebi, M. Bacterial and viral infections. In: Varki, A., Cummings, R.D., Esko, J.D., et al., editors. Essentials of Glycobiology [Internet]. 3rd edition. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press; 2015-2017. Chapter 42 (2017).
Chan, J., Watson, J.N., Lu, A., Cerda, V.C., Borgford, T.J., Bennet, A.J.: Bacterial and viral sialidases: contribution of the conserved active site glutamate to catalysis. Biochemistry.51, 433–441 (2012)
Roy, S., Honma, K., Douglas, C.W., Sharma, A., Stafford, G.P.: Role of sialidase in glycoprotein utilization by Tannerella forsythia. Microbiol.157, 3195–3202 (2011)
Suzuki, T.: Imaging of sialidase activity and its clinical application. Biol. Pharm. Bull.40, 2015–2023 (2017)
Zhang, Y., Albohy, A., Zou, Y., Smutova, V., Pshezhetsky, A.V., Cairo, C.W.: 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.56, 2948–2958 (2013)
Nan, X., Carubelli, I., Stamatos, N.M.: Sialidase expression in activated human T lymphocytes influences production of IFN-gamma. J. Leukoc. Biol. 81, 284–296 (2007)
Stamatos, N.M., Liang, F., Nan, X., Landry, K., Cross, A.S., Wang, L.X., Pshezhetsky, A.V.: Differential expression of endogenous sialidases of human monocytes during cellular differentiation into macrophages. FEBS J.272, 2545–2556 (2005)
Wang, D., Ozhegov, E., Wang, L., Zhou, A., Nie, H., Li, Y., Sun, X.-L.: Sialylation and desialylation dynamics of monocytes upon differentiation and polarization to macrophages. Glycoconjugate J.33, 725–733 (2016)
Corfield, A.P., Higa, H., Paulson, J.C., Schauer, R.: The specificity of viral and bacterial sialidases for alpha(2-3)- and alpha(2-6)-linked sialic acids in glycoproteins. Biochim. Biophys. Acta. 744, 121–126 (1983)
Yu, K., Tsai, Y.-T., Ariga, T., Yanagisawa, M.: Structures, biosynthesis, and functions of gangliosides - An overview. J. Oleo. Sci.60, 537–544 (2011)
Prokazova, N.V., Samovilova, N.N., Gracheva, E.V., Golovanova, N.K.: Ganglioside GM3 and its biological functions. Biochemistry (Moscow). 74, 235–249 (2009)
Wolf, A.A., Fujinaga, Y., Lencer, W.I.: Uncoupling of the cholera toxin-GM1 ganglioside receptor complex from endocytosis, retrograde Golgi trafficking, and downstream signal transduction by depletion of membrane cholesterol. J. Biol. Chem. 277, 16249–16256 (2002)
Ravindran, M.S., Tanner, L.B., Wenk, M.R.: Sialic acid linkage in glycosphingolipids is a molecular correlate for trafficking and delivery of extracellular cargo. Traffic. 14, 1182–1191 (2013)
Rodriguez, J.A., Piddini, E., Hasegawa, T., Miyagi, T., Dotti, C.G.: Plasma membrane ganglioside sialidase regulates axonal growth and regeneration in hippocampal neurons in culture. J. Neurosci.21, 8387–8395 (2001)
Kappagantula, S., Andrews, M.R., Cheah, M., Abad-Rodriguez, J., Dotti, C.G., Fawcett, J.W.: Neu3 sialidase-mediated ganglioside conversion is necessary for axon regeneration and is blocked in CNS axons. J. Neurosci.34, 2477–2492 (2014)
Pan, X, De Aragão, CBP, Velasco-Martin, JP, Priestman, DA, Wu, HY, Takahashi, K, Yamaguchi, K., Sturiale, L., Garozzo, D., Platt, F.M., Lamarche-Vane, N., Morales, C.R., Miyagi, T., Pshezhetsky, A.V.: Neuraminidases 3 and 4 regulate neuronal function by catabolizing brain gangliosides. FASEB J.31, 3467-3483(2017).
Berenson, C.S., Nawar, H.F., Yohe, H.C., Castle, S.A., Ashline, D.J., Reinhold, V.N., Hajishengallis, G., Connell, T.D.: Mammalian cell ganglioside-binding specificities of E. coli enterotoxins LT-IIb and variant LT-IIb(T13I). Glycobiology. 20, 41-54 (2010).
Shenoy, G.N., Loyall, J., Berenson, C.S., Kelleher Jr., R.J., Iyer, V., Balu-Iyer, S.V., Odunsi, K., Bankert, R.B.: Sialic acid-dependent inhibition of T cells by exosomal ganglioside GD3 in ovarian tumor microenvironments. J. Immunol.201, 3750–3758 (2018)
Tringali, C., Papini, N., Fusi, P., Croci, G., Borsani, G., Preti, A., Tortora, P., Tettamanti, G., Venerando, B., Monti, E.: Properties of recombinant human cytosolic sialidase HsNEU2. The enzyme hydrolyzes monomerically dispersed GM1 ganglioside molecules. J. Biol. Chem.279, 3169-79 (2004).
Chokhawala, H.A., Yu, H., Chen, X.: High-throughput substrate specificity studies of sialidases by using chemoenzymatically synthesized sialoside libraries. ChemBioChem.8, 194–201 (2007)
Kopitz, J., Sinz, K., Brossmer, R., Cantz, M.: Partial characterization and enrichment of a membrane-bound sialidase specific for gangliosides from human brain tissue. Eur. J. Biochem.248, 527–534 (1997)
Gatt, S., Gazit, B., Cestaro, B., Barenholz, Y.: Hydrolysis of gangliosides in micellar and liposomal dispersion by bacterial neuraminidases. Adv. Exp. Med. Biol.125, 137–146 (1980)
Venerando, B., Cestaro, B., Fiorilli, A., Ghidoni, R., Preti, A., Tettamanti, G.: Kinetics of Vibrio cholerae sialidase action on gangliosidic substrates at different supramolecular-organizational levels. Biochem J.203, 735–742 (1982)
Simons, K., Sampaio, J.L.: Membrane organization and lipid rafts. Cold Spring Harb Perspect. Biol.3, a004697 (2011)
Sanders, C.R., Mittendorf, K.F.: Tolerance to changes in membrane lipid composition as a selected trait of membrane proteins. Biochemistry.50, 7858–7867 (2011)
Saito, M., Sugano, K., Nagai, Y.: Action of Arthrobacter ureafaciens sialidase on sialoglycolipid substrates. Mode of action and highly specific recognition of the oligosaccharide moiety of ganglioside GM1. J. Biol. Chem.254, 7845-7854 (1979).
Hitika, T., Aritomi, K., Tanaka, N., Toyoda, H., Suzuki, A., Toida, T., Abe, T., Yanagawa, Y., Ishizuka, I.: Determination of N-acetyl- and N-glycolylneuraminic acids in gangliosides by combination of neuraminidase hydrolysis and fluorometric high-performance liquid chromatography using a GM3 derivative as an internal standard. Anal. Biochem.281, 193–201 (2000)
Stewart, J.C.: Colorimetric determination of phospholipids with ammonium ferrothiocyanate. Anal. Biochem.104, 10–14 (1980)
Murate, M., Abe, M., Kasahara, K., Iwabuchi, K., Umeda, M., Kobayashi, T.: Transbilayer distribution of lipids at nano scale. J. Cell Sci.128, 1627–1638 (2015)
McMahon, H.T., Boucrot, E.: Membrane curvature at a glance. J. Cell Sci.128, 1065–1070 (2015)
Wang, L., Wang, D., Zhou, X., Wu, L., Sun, X.-L.: Systematic investigation of quinoxaline derivatization of sialic acids and their quantitation applicability using high performance liquid chromatography. RSC Adv.4, 45797–45803 (2014)
van Meer, G.: Dynamic Transbilayer Lipid Asymmetry. Cold Spring Harb Perspect Biol.3, a004671 (2011)
Thomas, P.D., Poznansky, M.J.: Curvature and composition-dependent lipid asymmetry in phosphatidylcholine vesicles containing phosphatidylethanolamine and gangliosides. Biochim Biophys Acta. 978, 85–90 (1989)
Acknowledgments
This work was supported by Faculty Research Fund from the Center for Gene Regulation in Health and Disease (GRHD) and Faculty Research Development (FRD) Fund at Cleveland State University.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflicts of interest.
Ethical approval
This article does not contain any studies with human participants or animals performed by any of the authors.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Tomar, S., Sun, XL. Investigation of substrate specificity of sialidases with membrane mimetic glycoconjugates. Glycoconj J 37, 175–185 (2020). https://doi.org/10.1007/s10719-019-09895-x
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10719-019-09895-x