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Publications (10 of 20) Show all publications
Behren, S., Schorlemer, M., Schmidt, G., Westerlind, U. & Aktories, K. (2023). Antibodies directed against GalNAc- and GlcNAc-O-Tyrosine posttranslational modifications – a new tool for glycoproteomic detection. Chemistry - A European Journal, 29(29), Article ID e202300392.
Open this publication in new window or tab >>Antibodies directed against GalNAc- and GlcNAc-O-Tyrosine posttranslational modifications – a new tool for glycoproteomic detection
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2023 (English)In: Chemistry - A European Journal, ISSN 0947-6539, E-ISSN 1521-3765, Vol. 29, no 29, article id e202300392Article in journal (Refereed) Published
Abstract [en]

In the last decade, it was discovered that protein mucin-type O-glycosylation and O-GlcNAcylation modify Tyr residues besides the well explored Thr and Ser amino acids. Several glycoproteomic studies have identified α-GalNAc-O-Tyr modifications, and studies propose that β-GlcNAc-O-Tyr also exists as a new group of posttranslational modifications (PTMs). Specific bacterial toxins have further been identified to modify host GTPases with α-GlcNAc-O-Tyr to promote bacterial virulence. Despite being identified on numerous proteins, the biological roles, biosynthesis and expression of GalNAc- and GlcNAc-O-Tyr modifications are poorly understood. A major obstacle is the lack of tools to specifically detect and identify proteins containing these modifications. With this in mind, we prepared vaccine constructs and raised antibodies to enable selective detection of proteins carrying these new PTMs. The obtained polyclonal antibody sera were evaluated using ELISA and glycopeptide microarrays and were found to be highly selective for GlcNAc- and GalNAc-O-Tyr glycopeptides over the corresponding Ser- and Thr-modifications. For microarray analysis, synthetic GlcNAc- and GalNAc-O-Tyr Fmoc-amino acids were prepared and applied in Fmoc-SPPS to obtain an extensive O-glycopeptide library. After affinity purification, the antibodies were applied in western blot analysis and showed specific detection of α-GlcNAc-O-Tyr modified RhoA GTPase.

Place, publisher, year, edition, pages
Wiley-VCH Verlagsgesellschaft, 2023
Keywords
antibodies, vaccine, glycoproteomics, glycopeptides, glycosylation, PTM
National Category
Organic Chemistry
Research subject
Biorganic Chemistry; biological chemistry
Identifiers
urn:nbn:se:umu:diva-187279 (URN)10.1002/chem.202300392 (DOI)000970554600001 ()37052513 (PubMedID)2-s2.0-85152586008 (Scopus ID)
Note

Originally included in thesis in manuscript form. 

Available from: 2021-09-07 Created: 2021-09-07 Last updated: 2023-06-19Bibliographically approved
Behren, S., Yu, J., Pett, C., Schorlemer, M., Heine, V., Fischöder, T., . . . Westerlind, U. (2023). Fucose binding motifs on mucin core glycopeptides impact bacterial lectin recognition. Angewandte Chemie International Edition, 62(32), Article ID e202302437.
Open this publication in new window or tab >>Fucose binding motifs on mucin core glycopeptides impact bacterial lectin recognition
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2023 (English)In: Angewandte Chemie International Edition, ISSN 1433-7851, E-ISSN 1521-3773, Vol. 62, no 32, article id e202302437Article in journal (Refereed) Published
Abstract [en]

Mucin glycoproteins are essential components of the mucosal barrier, which protects the host from pathogens. Throughout evolution, bacteria have developed strategies to modulate and penetrate this barrier, and cause virulence by interacting with mucin O-glycans at the epithelial cell-surface. O-fucosylated glycan epitopes on mucins are key ligands of many bacterial lectins. Here, a chemoenzymatic synthesis strategy is described to prepare a library of fucosylated mucin core glycopeptides to enable studies of mucin-interacting and fucose-binding bacterial lectins. Glycan cores with biologically important Lewis and H-antigens were prepared decorating the peptide backbone at different sites and densities. The fucosylated mucin glycopeptides were applied in microarray binding studies to explore the importance of glycan core and peptide backbone presentation of these antigens in binding interactions with the P. aeruginosa lectin LecB and the C. difficile toxin A.

Keywords
Fucose, Glycopeptides, Glycosylation, Lectins, Microarrays
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:umu:diva-209298 (URN)10.1002/anie.202302437 (DOI)000989616900001 ()2-s2.0-85159456005 (Scopus ID)
Funder
Swedish Research Council, 2020-04160The Kempe Foundations
Available from: 2023-06-08 Created: 2023-06-08 Last updated: 2023-10-16Bibliographically approved
Gabba, A., Attariya, R., Behren, S., Pett, C., van der Horst, J. C., Yurugi, H., . . . Murphy, P. V. (2023). MUC1 glycopeptide vaccine modified with a GalNAc glycocluster targets the macrophage galactose c-type lectin on dendritic cells to elicit an improved humoral response. Journal of the American Chemical Society, 145(24), 13027-13037
Open this publication in new window or tab >>MUC1 glycopeptide vaccine modified with a GalNAc glycocluster targets the macrophage galactose c-type lectin on dendritic cells to elicit an improved humoral response
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2023 (English)In: Journal of the American Chemical Society, ISSN 0002-7863, E-ISSN 1520-5126, Vol. 145, no 24, p. 13027-13037Article in journal (Refereed) Published
Abstract [en]

Mucin expression and glycosylation patterns on cancer cells differ markedly from healthy cells. Mucin 1 (MUC1) is overexpressed in several solid tumors and presents high levels of aberrant, truncated O-glycans (e.g., Tn antigen). Dendritic cells (DCs) express lectins that bind to these tumor-associated carbohydrate antigens (TACAs) to modulate immune responses. Selectively targeting these receptors with synthetic TACAs is a promising strategy to develop anticancer vaccines and to overcome TACA tolerance. In this work, we prepared, via a solid phase peptide synthesis approach, a modular tripartite vaccine candidate, incorporating a high-affinity glycocluster based on a tetraphenylethylene scaffold, to target the macrophage galactose-type lectin (MGL) on antigen presenting cells. MGL is a C-type lectin receptor that binds Tn antigens and can route them to human leukocyte antigen class II or I, making it an attractive target for anticancer vaccines. Conjugation of the glycocluster to a library of MUC1 glycopeptides bearing the Tn antigen is shown to promote uptake and recognition of the TACA by DCs via MGL. In vivo testing revealed that immunization with the newly designed vaccine construct bearing the GalNAc glycocluster induced a higher titer of anti-Tn-MUC1 antibodies compared to the TACAs alone. Additionally, the antibodies obtained bind a library of tumor-associated saccharide structures on MUC1 and MUC1-positive breast cancer cells. Conjugation of a high-affinity ligand for MGL to tumor-associated MUC1 glycopeptide antigens has a synergistic impact on antibody production.

Place, publisher, year, edition, pages
American Chemical Society (ACS), 2023
National Category
Other Basic Medicine
Identifiers
urn:nbn:se:umu:diva-211840 (URN)10.1021/jacs.2c12843 (DOI)001008122000001 ()37279388 (PubMedID)2-s2.0-85162847018 (Scopus ID)
Funder
German Research Foundation (DFG), CRC 1066EU, Horizon 2020, ERC CoG SUPRAVACC819856The Kempe Foundations, JCK-1819.1The Kempe Foundations, JCK-1819.2
Available from: 2023-07-11 Created: 2023-07-11 Last updated: 2023-07-11Bibliographically approved
Behren, S. & Westerlind, U. (2023). Novel approaches to design glycan-based antibacterial inhibitors. European Journal of Organic Chemistry, 26(1), Article ID e202200795.
Open this publication in new window or tab >>Novel approaches to design glycan-based antibacterial inhibitors
2023 (English)In: European Journal of Organic Chemistry, ISSN 1434-193X, E-ISSN 1099-0690, Vol. 26, no 1, article id e202200795Article, review/survey (Refereed) Published
Abstract [en]

The interactions between bacterial lectins and carbohydrates on the host cell surface can mediate bacterial adhesion, invasion, and immune evasion. Multivalency plays a key role in these binding events. However, additional molecular mechanisms greatly impact multivalent binding recognition. To develop specific and effective bacterial inhibitors, a deeper understanding of the complex underlying mechanisms of bacterial adhesion processes is necessary. By interfering with bacterial adhesion, synthetic multivalent glycoconjugates do not only have the potential to improve or replace antibiotic treatments, but also represent useful tools to study carbohydrate-pathogen interactions. In this review, we highlight a few recent advances in the synthesis and application of synthetic glycan-based scaffolds to uncover the nature of glycan-bacteria interactions and to design efficient bacterial inhibitors.

Place, publisher, year, edition, pages
John Wiley & Sons, 2023
Keywords
Bacteria, Glycoconjugates, Inhibitors, Lectin, Multivalency
National Category
Organic Chemistry
Identifiers
urn:nbn:se:umu:diva-200569 (URN)10.1002/ejoc.202200795 (DOI)000868847200001 ()2-s2.0-85139931624 (Scopus ID)
Available from: 2022-10-31 Created: 2022-10-31 Last updated: 2023-01-11Bibliographically approved
Jiang, S., Wang, T., Behren, S., Westerlind, U., Gawlitza, K., Persson, J. L. & Rurack, K. (2022). Sialyl-tn antigen-imprinted dual fluorescent core-shell nanoparticles for ratiometric Sialyl-Tn antigen detection and dual-color labeling of cancer cells. ACS Applied Nano Materials, 5(12), 17592-17605
Open this publication in new window or tab >>Sialyl-tn antigen-imprinted dual fluorescent core-shell nanoparticles for ratiometric Sialyl-Tn antigen detection and dual-color labeling of cancer cells
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2022 (English)In: ACS Applied Nano Materials, E-ISSN 2574-0970, Vol. 5, no 12, p. 17592-17605Article in journal (Refereed) Published
Abstract [en]

Sialyl-Tn (STn or sialyl-Thomsen-nouveau) is a carbohydrate antigen expressed by more than 80% of human carcinomas. We here report a strategy for ratiometric STn detection and dual-color cancer cell labeling, particularly, by molecularly imprinted polymers (MIPs). Imprinting was based on spectroscopic studies of a urea-containing green-fluorescent monomer 1 and STn-Thr-Na (sodium salt of Neu5Acα2-6GalNAcα-O-Thr). A few-nanometer-thin green-fluorescent polymer shell, in which STn-Thr-Na was imprinted with 1, other comonomers, and a cross-linker, was synthesized from the surface of red-emissive carbon nanodot (R-CND)-doped silica nanoparticles, resulting in dual fluorescent STn-MIPs. Dual-color labeling of cancer cells was achieved since both red and green emissions were detected in two separate channels of the microscope and an improved accuracy was obtained in comparison with single-signal MIPs. The flow cytometric cell analysis showed that the binding of STn-MIPs was significantly higher (p < 0.001) than that of non-imprinted polymer (NIP) control particles within the same cell line, allowing to distinguish populations. Based on the modularity of the luminescent core-fluorescent MIP shell architecture, the concept can be transferred in a straightforward manner to other target analytes.

Place, publisher, year, edition, pages
American Chemical Society (ACS), 2022
Keywords
cancer, core-shell particles, dual-color labeling, glycan, molecular imprinting
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:umu:diva-201480 (URN)10.1021/acsanm.2c03252 (DOI)000886696300001 ()2-s2.0-85142609638 (Scopus ID)
Funder
EU, Horizon 2020, 721297
Available from: 2022-12-06 Created: 2022-12-06 Last updated: 2023-01-11Bibliographically approved
Li, C., Palma, A. S., Zhang, P., Zhang, Y., Gao, C., Silva, L. M., . . . Chai, W. (2021). Noncovalent microarrays from synthetic amino-terminating glycans: Implications in expanding glycan microarray diversity and platform comparison. Glycobiology, 31(8), 931-946
Open this publication in new window or tab >>Noncovalent microarrays from synthetic amino-terminating glycans: Implications in expanding glycan microarray diversity and platform comparison
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2021 (English)In: Glycobiology, ISSN 0959-6658, E-ISSN 1460-2423, Vol. 31, no 8, p. 931-946Article in journal (Refereed) Published
Abstract [en]

Glycan microarrays have played important roles in detection and specificity assignment of glycan recognition by proteins. However, the size and diversity of glycan libraries in current microarray systems are small compared to estimated glycomes, and these may lead to missed detection or incomplete assignment. For microarray construction, covalent and noncovalent immobilization are the two types of methods used, but a direct comparison of results from the two platforms is required. Here we develop a chemical strategy to prepare lipid-linked probes from both naturally derived aldehyde-terminating and synthetic amino-terminating glycans that addresses the two aspects: expansion of sequence-defined glycan libraries and comparison of the two platforms. We demonstrate the specific recognition by plant and mammalian lectins, carbohydrate-binding modules and antibodies and the overall similarities from the two platforms. Our results provide new knowledge on unique glycan-binding specificities for the immune receptor Dectin-1 toward beta-glucans and the interaction of rotavirus P[19] adhesive protein with mucin O-glycan cores.

Place, publisher, year, edition, pages
Oxford University Press, 2021
Keywords
Dectin-1, glycan micorarray, oligosaccharide microarray, neoglycolipid, rotavirus
National Category
Microbiology in the medical area Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:umu:diva-188092 (URN)10.1093/glycob/cwab037 (DOI)000697400400006 ()33978739 (PubMedID)2-s2.0-85116345286 (Scopus ID)
Available from: 2021-10-13 Created: 2021-10-13 Last updated: 2021-10-18Bibliographically approved
Wu, X., McFall-Boegeman, H., Rashidijahanabad, Z., Liu, K., Pett, C., Yu, J., . . . Huang, X. (2021). Synthesis and immunological evaluation of the unnatural β-linked mucin-1 Thomsen-Friedenreich conjugate. Organic and biomolecular chemistry, 19(11), 2448-2455
Open this publication in new window or tab >>Synthesis and immunological evaluation of the unnatural β-linked mucin-1 Thomsen-Friedenreich conjugate
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2021 (English)In: Organic and biomolecular chemistry, ISSN 1477-0520, E-ISSN 1477-0539, Vol. 19, no 11, p. 2448-2455Article in journal (Refereed) Published
Abstract [en]

MUC1 glycopeptides are attractive antigens for anti-cancer vaccine development. One potential drawback in using the native MUC1 glycopeptide for vaccine design is the instability of theO-glycosyl linkage between the glycan and the peptide backbone to glycosidase. To overcome this challenge, a MUC1 glycopeptide mimic has been synthesized with the galactose-galactosamine disaccharide linked with threonine (Thomsen-Friedenreich or Tf antigen) through an unnatural β-glycosyl bond. The resulting MUC1-β-Tf had a much-enhanced stability toward a glycosidase capable of cleaving the glycan from the corresponding MUC1 glycopeptide with the natural α-Tf linkage. The MUC1-β-Tf was subsequently conjugated with a powerful carrier bacteriophage Qβ. The conjugate induced high levels of IgG antibodies in clinically relevant human MUC1 transgenic mice, which cross-recognized not only the natural MUC1-α-Tf glycopeptide but also MUC1 expressing tumor cells, supporting the notion that a simple switch of the stereochemistry of the glycan/peptide linkage can be a strategy for anti-cancer vaccine epitope design for glycopeptides.

Place, publisher, year, edition, pages
Royal Society of Chemistry, 2021
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
urn:nbn:se:umu:diva-182097 (URN)10.1039/d1ob00007a (DOI)000632575900009 ()2-s2.0-85103271975 (Scopus ID)
Funder
NIH (National Institute of Health), R01 CA225105
Available from: 2021-04-15 Created: 2021-04-15 Last updated: 2023-09-05Bibliographically approved
Behren, S. & Westerlind, U. (2019). Glycopeptides and -Mimetics to Detect, Monitor and Inhibit Bacterial and Viral Infections: Recent Advances and Perspectives. Molecules, 24(6), Article ID 1004.
Open this publication in new window or tab >>Glycopeptides and -Mimetics to Detect, Monitor and Inhibit Bacterial and Viral Infections: Recent Advances and Perspectives
2019 (English)In: Molecules, ISSN 1431-5157, E-ISSN 1420-3049, Vol. 24, no 6, article id 1004Article, review/survey (Refereed) Published
Abstract [en]

The initial contact of pathogens with host cells is usually mediated by their adhesion to glycan structures present on the cell surface in order to enable infection. Furthermore, glycans play important roles in the modulation of the host immune responses to infection. Understanding the carbohydrate-pathogen interactions are of importance for the development of novel and efficient strategies to either prevent, or interfere with pathogenic infection. Synthetic glycopeptides and mimetics thereof are capable of imitating the multivalent display of carbohydrates at the cell surface, which have become an important objective of research over the last decade. Glycopeptide based constructs may function as vaccines or anti-adhesive agents that interfere with the ability of pathogens to adhere to the host cell glycans and thus possess the potential to improve or replace treatments that suffer from resistance. Additionally, synthetic glycopeptides are used as tools for epitope mapping of antibodies directed against structures present on various pathogens and have become important to improve serodiagnostic methods and to develop novel epitope-based vaccines. This review will provide an overview of the most recent advances in the synthesis and application of glycopeptides and glycopeptide mimetics exhibiting a peptide-like backbone in glycobiology.

Place, publisher, year, edition, pages
MDPI, 2019
Keywords
glycopeptides, glycopeptide mimetics, host-pathogen interactions, serodiagnostics
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:umu:diva-157846 (URN)10.3390/molecules24061004 (DOI)000465503800004 ()30871155 (PubMedID)2-s2.0-85062978043 (Scopus ID)
Available from: 2019-04-04 Created: 2019-04-04 Last updated: 2023-08-28Bibliographically approved
Broman, K., Mårell-Olsson, E., Johnels, D., Andersson, C. D., Chorell, E., Westerlind, U., . . . Norrby, M. (2019). Spatial Ability in Organic Chemistry: Can Virtual and Augmented Reality be Valuable?. In: 7:e Utvecklingskonferensen för Sveriges ingenjörsutbildningar: . Paper presented at 7:e Utvecklingskonferensen för Sveriges ingenjörsutbildningar 2019, Luleå tekniska universitet, Luleå, 27–28 november, 2019. Luleå tekniska universitet
Open this publication in new window or tab >>Spatial Ability in Organic Chemistry: Can Virtual and Augmented Reality be Valuable?
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2019 (English)In: 7:e Utvecklingskonferensen för Sveriges ingenjörsutbildningar, Luleå tekniska universitet , 2019Conference paper, Published paper (Other academic)
Abstract [en]

In this paper, the roles of digital technologies as Virtual Reality (VR), and Augmented Reality (AR), are discussed to explore how biotechnology engineering students develop their spatial ability in organic chemistry. We have, through stereochemistry workshops, followed how students, in specific, visualise and rotate molecular representations and how the use of digital tools influences the students’ interest.

Place, publisher, year, edition, pages
Luleå tekniska universitet, 2019
Keywords
Spatial ability, Visualisation, Virtual reality, Augmented reality, Organic chemistry, Lewis structures, Biotechnology engineering students
National Category
Didactics
Research subject
didactics of chemistry
Identifiers
urn:nbn:se:umu:diva-165507 (URN)
Conference
7:e Utvecklingskonferensen för Sveriges ingenjörsutbildningar 2019, Luleå tekniska universitet, Luleå, 27–28 november, 2019
Available from: 2019-11-28 Created: 2019-11-28 Last updated: 2022-12-02Bibliographically approved
Wu, X., McKay, C., Pett, C., Yu, J., Schorlemer, M., Ramadan, S., . . . Huang, X. (2019). Synthesis and Immunological Evaluation of Disaccharide Bearing MUC-1 Glycopeptide Conjugates with Virus-like Particles. ACS Chemical Biology, 14(10), 2176-2184
Open this publication in new window or tab >>Synthesis and Immunological Evaluation of Disaccharide Bearing MUC-1 Glycopeptide Conjugates with Virus-like Particles
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2019 (English)In: ACS Chemical Biology, ISSN 1554-8929, E-ISSN 1554-8937, Vol. 14, no 10, p. 2176-2184Article in journal (Refereed) Published
Abstract [en]

Mucin-1 (MUC1) is a highly attractive antigenic target for anticancer vaccines. Naturally existing MUC1 can contain multiple types of O-linked glycans, including the Thomsen–Friedenreich (Tf) antigen and the Sialyl Thomsen-nouveau (STn) antigen. In order to target these antigens as potential anticancer vaccines, MUC1 glycopeptides SAPDT*RPAP (T* is the glycosylation site) bearing the Tf and the STn antigen, respectively, have been synthesized. The bacteriophage Qβ carrier is a powerful carrier for antigen delivery. The conjugates of MUC1-Tf and -STn glycopeptides with Qβ were utilized to immunize immune-tolerant human MUC1 transgenic (MUC1.Tg) mice, which elicited superior levels of anti-MUC1 IgG antibodies with titers reaching over 2 million units. The IgG antibodies recognized a wide range of MUC1 glycopeptides bearing diverse glycans. Antibodies induced by Qβ-MUC1-Tf showed strongest binding, with MUC1-expressing melanoma B16-MUC1 cells, and effectively killed these cells in vitro. Vaccination with Qβ-MUC1-Tf first followed by tumor challenge in a lung metastasis model showed significant reductions of the number of tumor foci in the lungs of immunized mice as compared to those in control mice. This was the first time that a MUC1-Tf-based vaccine has shown in vivo efficacy in a tumor model. As such, Qβ-MUC1 glycopeptide conjugates have great potential as anticancer vaccines.

Place, publisher, year, edition, pages
American Chemical Society (ACS), 2019
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
urn:nbn:se:umu:diva-165120 (URN)10.1021/acschembio.9b00381 (DOI)000492118500010 ()31498587 (PubMedID)2-s2.0-85073024998 (Scopus ID)
Available from: 2019-11-11 Created: 2019-11-11 Last updated: 2023-03-24Bibliographically approved
Organisations
Identifiers
ORCID iD: ORCID iD iconorcid.org/0000-0002-4841-6238

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