Glyco-Enzyme Repository

Repository of Expression Constructs for Glycosylation Enzymes

Gateway® entry clones and mammalian, baculovirus and bacterial expression constructs for glycosyltransferases, glycoside hydrolases and glycan-modifying enzymes

Home/Publications

Publications

51 publications that used the expression technologies, constructs or proteins produced by the Repository, grouped as on the original site. Several entries were listed as in press when the legacy page was last updated; their volume and page numbers are given where the citation recorded them.

Citation. If you use Repository constructs or proteins in your research, please cite this website (glycoenzymes.ccrc.uga.edu) and the grant support for the Repository (National Institutes of Health grants P41GM103390 and P01GM107012) in any publications.

General methods of mammalian cell expression

  1. Moremen, K.W., Ramiah, A., Stuart, M., Steel, J., Meng, L., Forouhar, F., Moniz, H.A., Gahlay, G., Gao, Z., Chapla, D., Wang, S., Yang, J.-Y., Prabahkar, P.K., Johnson, R., dela Rosa, M., Geisler, C., Nairn, A.V., Wu, S.-C., Tong, L., Gilbert, H.J., LaBaer, J. and Jarvis, D.L. (2018) Expression system for structural and functional studies of human glycosylation enzymes. Nature Chem. Biol. 14, 156–162. [PMID 29251719; PMC5774587]
  2. Subedi, G.P., Moniz, H., Johnson, R.W., Moremen, K.W. and Barb, A.W. (2015) High yield expression of recombinant human proteins with the transient transfection of HEK293 cells in suspension. J. Vis. Exp. 106:e53568. [PMID 26779721; PMC4780855]

Characterisation of glycosylation enzymes produced in the Repository

  1. Thieker, D., Xu, Y., Chapla, D., Nora, C., Qiu, H., Felix, T., Wang, L., Moremen, K.W., Liu, J., Esko, J. and Woods, R.J. (2018) Downstream products are potent inhibitors of the heparan sulfate 2-O-sulfotransferase. Scientific Reports 8, 11823. [PMID 30087361; PMC6081452]
  2. Jensen, J.K., Busse-Wicher, M., Poulsen, C.P., Fangel, J.U., Smith, P.J., Yang, J.-Y., Peña, M.-J., Dinesen, M.H., Martens, H.J., Melkonian, M., Wong, G.K., Moremen, K.W., Wilkerson, C.G., Scheller, H.V., Dupree, P., Ulvskov, P., Urbanowicz, B.R. and Harholt, J. (2018) Identification of an algal xylan synthase KfXYS1 proves that there is functional orthology between algal and plant cell wall biosynthesis. New Phytologist 218, 1049–1060. [PMID 29460505; PMC5902652]
  3. Hanes, M.S., Moremen, K.W. and Cummings, R.D. (2017) Biochemical characterization of functional domains of the chaperone Cosmc. PLOS One 12:e0180242. [PMID 28665962; PMC5493369]
  4. Urbanowicz, B.R., Bharadwaj, V.S., Alahuhta, M., Peña, M.J., Lunin, V.V., Bomble, Y.J., Wang, S., Yang, J.-Y., Tuomivaara, S., Himmel, M.E., Moremen, K.W., York, W.S. and Crowley, M.F. (2017) Structural, mutagenic and in silico studies of xyloglucan fucosylation in Arabidopsis thaliana suggest a water-mediated mechanism. Plant J. 91, 931–949. [PMID 28670741; PMC5735850]
  5. Sheikh, M.O., Halmo, S.M., Patel, S., Middleton, D., Takeuchi, H., Schafer, C.M., West, C.M., Haltiwanger, R.S., Avci, F.Y., Moremen, K.W. and Wells, L. (2017) Rapid screening of sugar-nucleotide donor specificities of putative glycosyltransferases. Glycobiology 27, 206–212. [PMID 28177478; PMC5789813]
  6. Halmo, S.M., Singh, D., Patel, S., Wang, S., Edlin, E., Boons, G.J., Moremen, K.W., Live, D.H. and Wells, L. (2017) Protein O-linked mannose β-1,4-N-acetylglucosaminyltransferase 2 (POMGNT2) is a gatekeeper enzyme for functional glycosylation of α-dystroglycan. J. Biol. Chem. 292, 2101–2109. [PMID 27932460; PMC5313085]
  7. Li, T., Huang, M., Liu, L., Wang, S., Moremen, K.W. and Boons, G.J. (2016) Divergent chemoenzymatic synthesis of asymmetrical core-fucosylated and core-unmodified N-glycans. Chemistry 22, 18742–18746. [PMID 27798819; PMC5442444]
  8. Calderon, A.D., Liu, Y., Li, X., Wang, X., Chen, X., Li, L. and Wang, P.G. (2016) Substrate specificity of FUT8 and chemoenzymatic synthesis of core-fucosylated asymmetric N-glycans. Org. Biomol. Chem. 14, 4027–4031. [PMID 27080952; PMC4852481]
  9. Praissman, J.L., Willer, T., Sheikh, M.O., Toi, A., Chitayat, D., Lin, Y.Y., Lee, H., Stalnaker, S., Wang, S., Prabhakar, P., Nelson, S.F., Stemple, D.L., Moore, S.A., Moremen, K.W., Campbell, K.P. and Wells, L. (2016) The functional O-mannose glycan on α-dystroglycan contains a phospho-ribitol primed for matriglycan addition. eLife 5:e14473. [PMID 27130732; PMC4924997]
  10. Revoredo, L., Wang, S., Bennett, E.P., Clausen, H., Moremen, K.W., Jarvis, D.L., Ten Hagen, K., Tabak, L.A. and Gerken, T.A. (2016) Mucin-type O-glycosylation is controlled by short- and long-range glycopeptide substrate recognition that varies among members of the polypeptide GalNAc transferase (ppGalNAc-T) family. Glycobiology 26, 360–376. [PMID 26610890; PMC4767052]
  11. Wu, L., Viola, C.M., Brzozowski, A.M. and Davies, G.J. (2015) Structural characterization of human heparanase reveals insights into substrate recognition. Nat. Struct. Mol. Biol. 22, 1016–1023. [PMID 26575439; PMC5008439]
  12. Czuchry, D., Desormeaux, P., Stuart, M., Jarvis, D.L., Matta, K., Szarek, W.A. and Brockhausen, I. (2015) Synthesis of the sialyl-T antigen: biochemical characterization of a novel α2,3-sialyltransferase WbwA from pathogenic Escherichia coli serotype O104 and comparison to human ST3GAL1. J. Bacteriol. 197, 3760–3768. [PMC4652054]
  13. Praissman, J.L., Live, D.H., Wang, S., Ramiah, A., Moremen, K.W. and Wells, L. (2014) B4GAT1 is the priming enzyme for the LARGE-dependent functional glycosylation of α-dystroglycan. eLife 3, e03943. [PMID 25279697; PMC4227051]
  14. Gao, Y., Aryal, R.P., Ju, T., Cummings, R.D., Gahlay, G., Jarvis, D.L., Matta, K.L., Vlahakis, J.Z., Szarek, W.A. and Brockhausen, I. (2013) Acceptor specificities and selective inhibition of recombinant human Gal- and GlcNAc-transferases that synthesize core structures 1, 2, 3 and 4 of O-glycans. Biochim. Biophys. Acta 1830, 4274–4281. [PMC4002258]
  15. Gerken, T., Revoredo, L., Thome, J.J.C., Tabak, L.A., Vester-Christensen, M.B., Clausen, H., Gahlay, G.K., Jarvis, D.L., Johnson, R.Y., Moniz, H.A. and Moremen, K.W. (2013) The lectin domain of the polypeptide GalNAc transferase family of glycosyltransferases (ppGalNAc Ts) acts as a switch directing glycopeptide substrate glycosylation in an N- or C-direction, further controlling mucin-type O-glycosylation. J. Biol. Chem. 288, 19900–19914. [PMID 23689369; PMC3707691]
  16. Liu, S., Meng, L., Moremen, K.W. and Prestegard, J.H. (2009) NMR structural characterization of substrates bound to the α-2,6-sialyltransferase, ST6Gal-I. Biochemistry 48, 11211–11219. [PMID 19845399; PMC2790006]
  17. Macnaughtan, M.A., Tian, F., Liu, S., Meng, L., Park, S., Azadi, P., Moremen, K.W. and Prestegard, J.H. (2008) 13C-sialic acid labeling of glycans on glycoproteins using ST6Gal-I. J. Am. Chem. Soc. 130, 11864–11865. [PMID 18700760; PMC2640832]

Structure determination of glycosylation enzymes produced in the Repository

  1. Kadirvelraj, R., Yang, J.-Y., Sanders, J.H., Liu, L., Ramiah, A., Prabhakar, P.K., Boons, G.-J., Wood, Z.A. and Moremen, K.W. (2018) Human N-acetylglucosaminyltransferase II substrate recognition uses a modular architecture that includes a convergent exosite. Proc. Natl. Acad. Sci. USA 115, 4637–4642. [PMID 29666272; PMC5939069]
  2. Xiang, Y., Karaveg, K. and Moremen, K.W. (2016) Substrate recognition and catalysis by GH47 α-mannosidases involved in Asn-linked glycan maturation in the mammalian secretory pathway. Proc. Natl. Acad. Sci. USA 113, E7890–E7899. [PMID 27856750; PMC5150396]
  3. Zhang, C., Zhang, T., Zou, J., Miller, C.J., Gorkhali, R., Yang, J.-Y., Schilmiller, A., Wang, S., Huang, K., Brown, E.M., Moremen, K.W., Hu, J. and Yang, J.J. (2016) Structural basis for regulation of human calcium-sensing receptor by magnesium ions and an unexpected tryptophan derivative co-agonist. Science Advances 2:e1600241. [PMID 27386547; PMC4928972]
  4. Meng, L., Forouhar, F., Thieker, D., Gao, Z., Ramiah, A., Moniz, H., Xiang, Y., Seetharaman, J., Milaninia, S., Su, M., Bridger, R., Veillon, L., Azadi, P., Kornhaber, G., Wells, L., Montelione, G.T., Woods, R.J., Tong, L. and Moremen, K.W. (2013) Enzymatic basis for N-glycan sialylation: structure of rat α2,6-sialyltransferase (ST6GAL1) reveals conserved and unique features for glycan sialylation. J. Biol. Chem. 288, 34680–34698. [PMID 24155237; PMC3843080]

Use of Repository enzymes for glycan synthesis

  1. Liu, L., Prudden, A.R., Capicciotti, C.J., Bosman, G.P., Yang, J.Y., Chapla, D.G., Moremen, K.W. and Boons, G.J. (2018) Streamlining the chemoenzymatic synthesis of complex N-glycans by a stop-and-go strategy. Nature Chem.
  2. Epp, A., Hobusch, J., Bartsch, Y., Eschweiler, S., Möbs, C., Hall, A., Morris, S.C., Petzold, D., Lilienthal, G.-M., Engellenner, C., Bitterling, J., Petry, J., Rahmöller, R., Collin, M., Moremen, K.W., Strait, R.T., Blanchard, V., Petersen, A., Gemoll, T., Habermann, J.K., Petersen, F., Nandy, A., Kahlert, H., Hertl, M., Pfützner, W., Jappe, U., Finkelman, F.D. and Ehlers, M. (2018) Sialylation of IgG antibodies inhibits IgG-mediated allergic reactions. J. Allergy Clin. Immunol. 141, 399–402. [PMID 28728998; PMC5758435]
  3. Benedetti, E., Pučić-Baković, M., Keser, T., Wahl, A., Hassinen, H., Trbojević Akmačić, I., Vila, M., Razdorov, G., Štambuk, J., Klarić, L., Ugrina, I., Selman, M., Wuhrer, M., Rudan, I., Polasek, O., Hayward, C., Grallert, H., Strauch, K., Peters, A., Meitinger, T., Gieger, C., Yang, J.-Y., Liu, L., Boons, G.-J., Kellokumpu, S., Moremen, K.W., Bovin, N., Theis, F.J., Lauc, G. and Krumsiek, J. (2017) Network inference from glycoproteomics data reveals new reactions in the IgG glycosylation pathway. Nat. Commun. 8, 1483. [PMID 29440641; PMC5811429]
  4. Prudden, A.R., Liu, L., Capicciotti, C.J., Wolfert, M.A., Wang, S., Gao, Z., Meng, L., Moremen, K.W. and Boons, G.-J. (2017) Synthesis of asymmetrical multi-antennary human milk oligosaccharides. Proc. Natl. Acad. Sci. USA 114, 6954–6959. [PMID 28630345; PMC5502611]
  5. Bello, C., Wang, S., Meng, L., Moremen, K.W. and Becker, C.F.W. (2015) A PEGylated photocleavable auxiliary mediates the site-specific glycosylation and native chemical ligation of peptides. Angew. Chem. Int. Ed. Engl. 54, 7711–7715. [PMID 25980981; PMC4524672]

Use of Repository enzymes for cell surface remodelling

  1. Jiang, H., López-Aguilar, A., Meng, L., Gao, Z., Liu, Y., Tian, X., Yu, G., Ovryn, B., Moremen, K.W. and Wu, P. (2018) Modulating cell-surface receptor signaling and ion channel functions by in situ glycan editing. Angew. Chem. Int. Ed. Engl. 57, 967–971. [PMID 29292859; PMC5779621]
  2. López-Aguilar, A., Meng, L., Hou, X., Li, W., Moremen, K.W. and Wu, P. (2018) Sialyltransferase-based chemoenzymatic histology for the detection of N- and O-glycans. Bioconjugate Chem. 29, 1231–1239. [PMID 29569918; PMC5972038]
  3. Briard, J.G., Jiang, H., Moremen, K.W., Macauley, M.S. and Wu, P. (2018) Cell-based glycan arrays for probing glycan–glycan binding protein interactions. Nature Commun. 9, 880. [PMID 29491407; PMC5830402]
  4. Rouhanifard, S.H., López-Aguilar, A., Meng, L., Izumori, K., Moremen, K.W. and Wu, P. (2018) Engineered glycocalyx regulates stem cell proliferation in crypt organoids. Cell Chem. Biol. 25, 1–8. [PMID 29429899; PMC5910180]
  5. Yu, S.H., Zhao, P., Sun, T., Gao, Z., Moremen, K.W., Boons, G.J., Wells, L. and Steet, R. (2016) Selective exo-enzymatic labeling detects increased cell surface sialoglycoprotein expression upon megakaryocytic differentiation. J. Biol. Chem. 291, 3982–3989. [PMID 26733198; PMC4759176]
  6. Mbua, N.E., Li, X., Flanagan-Steet, H.R., Meng, L., Moremen, K.W., Wolfert, M.A., Steet, R. and Boons, G.-J. (2013) Selective exo-enzymatic labeling (SEEL) of N-glycans of living cells by recombinant ST6Gal I. Angew. Chem. 52, 13012–13015. [PMID 24129959; PMC3869382]

Proteins produced using the Repository mammalian expression platform

  1. Moure, M.J., Eletsky, A., Gao, Q., Morris, L., Yang, J.Y., Chapla, D., Zhao, Y., Zong, C., Boons, G.J., Moremen, K.W. and Prestegard, J.H. (2018) Paramagnetic tags for glycosylation sites in glycoproteins: structural constraints on heparan sulfate binding to Robo1. ACS Chem. Biol. [PMID 30063822; PMC6161356]
  2. Amos, R.A., Pattathil, S., Yang, J.Y., Atmodjo, M.A., Urbanowicz, B.R., Moremen, K.W. and Mohnen, D. (2018) A two-phase model for the non-processive biosynthesis of homogalacturonan polysaccharides by the GAUT1:GAUT7 complex. J. Biol. Chem. [PMID 30327429]
  3. Muchero, W., Sondreli, K.L., Chen, J.-G., Urbanowicz, B.R., Zhang, J., Singan, V., Yang, Y., Brueggeman, R.S., Franco-Coronado, J., Abraham, N., Yang, J.-Y., Moremen, K.W., Weisberg, A.J., Chang, J.H., Lindquist, E., Berry, K., Ranjan, P., Jawdy, S., Schmutz, J., Tuskan, G.A. and LeBoldus, J.M. (2018) Association mapping, transcriptomics and transient expression identify candidate genes mediating plant–pathogen interactions in a tree. Proc. Natl. Acad. Sci. USA. [PMID 30337484]
  4. Voiniciuc, C., Engle, K., Günl, M., Dieluweit, S., Schmidt, M.H.-W., Yang, J.-Y., Moremen, K.W., Mohnen, D. and Usadel, B. (2018) Identification of key enzymes for pectin synthesis in Arabidopsis seed mucilage. Plant Physiol. [PMID 30228108]
  5. Gao, Q., Yang, J.Y., Moremen, K.W., Flanagan, J.G. and Prestegard, J.H. (2018) Structural characterization of a heparan sulfate pentamer interacting with LAR-Ig1-2. Biochemistry 57, 2189–2199. [PMID 29570275; PMC6015444]
  6. Zhao, Y., Yang, J.Y., Thieker, D.F., Xu, Y., Zong, C., Boons, G.-J., Liu, J., Woods, R.J., Moremen, K.W. and Amster, I.J. (2018) Traveling wave ion mobility spectrometry (TWIMS) study of the Robo1–heparan sulfate interaction. J. Am. Soc. Mass Spectrom. 29, 1153–1165. [PMID 29520710; PMC6004239]
  7. Gao, Q., Chen, C.Y., Zong, C., Wang, S., Morris, L.C., Boons, G.-J., Moremen, K.W. and Prestegard, J.H. (2016) Structural aspects of heparan sulfate binding to Robo1-D12. ACS Chemical Biology.
  8. Zhuo, Y., Yang, J.-Y., Moremen, K.W. and Prestegard, J. (2016) Glycosylation alters dimerization properties of a cell-surface signaling protein, CEACAM1. J. Biol. Chem.
  9. Li, Z., Moniz, H.A., Wang, S., Ramiah, A., Zhang, F., Moremen, K.W., Linhardt, R.J. and Sharp, J.S. (2015) High structural resolution hydroxyl radical protein footprinting reveals an extended Robo1–heparin binding interface. J. Biol. Chem. 290, 10729–10740. [PMID 25752613; PMC4409239]
  10. Zhang, C., Zhuo, Y., Moniz, H.A., Wang, S., Moremen, K.W., Prestegard, J.H., Brown, E. and Yang, J.J. (2014) Direct determination of multiple ligand interactions with the extracellular domain of the calcium-sensing receptor. J. Biol. Chem. 289, 33529–33542. [PMID 25305020; PMC4246106]
  11. Urbanowicz, B., Peña, M.J., Moniz, H.A., Moremen, K.W. and York, W.S. (2014) Two Arabidopsis proteins synthesize acetylated xylan in vitro. Plant J. 80, 197–206. [PMID 25141999; PMC4184958]
  12. Zhang, F., Moniz, H.A., Walcott, B., Moremen, K.W., Wang, L. and Linhardt, R.J. (2014) Probing the impact of GFP tagging on Robo1–heparin interaction. Glycoconj. J. 31, 299–307. [PMID 24748467; PMC4118743]
  13. Prestegard, J.H., Agard, D.A., Moremen, K.W., Lavery, L.A., Morris, L.C. and Pederson, K. (2014) Sparse labeling of proteins: structural characterization from long range constraints. J. Magn. Reson. 241, 32–40. [PMID 24656078; PMC3964372]
  14. Zhang, F., Moniz, H., Walcott, B., Moremen, K., Linhardt, R.J. and Wang, L. (2013) Characterization of the interaction between Robo1 and heparin and other glycosaminoglycans. Biochimie 95, 2345–2353. [PMID 23994753; PMC3871176]
  15. Lamanna, W.C., Lawrence, R., Sarrazin, S., Lameda-Diaz, C., Gordts, P.L.S.M., Moremen, K.W. and Esko, J.D. (2012) A genetic model of substrate reduction therapy for mucopolysaccharidosis. J. Biol. Chem. 287, 36283–36290. [PMID 22952226; PMC3476295]
  16. Barb, A.W., Meng, L., Gao, Z., Johnson, R.W., Moremen, K.W. and Prestegard, J.H. (2012) NMR characterization of immunoglobulin G Fc glycan motion on enzymatic sialylation. Biochemistry 51, 4618–4626. [PMID 22574931; PMC3447994]

Reviews

  1. Moremen, K.W., Tiemeyer, M. and Nairn, A.V. (2012) Vertebrate protein glycosylation: diversity, synthesis and function. Nat. Rev. Mol. Cell Biol. 13, 448–462. [PMID 22722607; PMC3934011]