TY - JOUR
T1 - (Automated) Synthesis of Well-defined Staphylococcus Aureus Wall Teichoic Acid Fragments
AU - Ali, Sara
AU - Hendriks, Astrid
AU - van Dalen, Rob
AU - Bruyning, Thomas
AU - Meeuwenoord, Nico
AU - Overkleeft, Herman S.
AU - Filippov, Dmitri V.
AU - van der Marel, Gijs A.
AU - van Sorge, Nina M.
AU - Codée, Jeroen D. C.
N1 - Funding Information: Simone Nicolardi and Fabrizio Chiodo for MALDI measurements. Carla de Haas and Piet Aerts for technical assistance with recombinant antibody production. This work was supported by Vidi (91713303) and Vici (09150181910001) grants from the Netherlands Organization for Health Research and Development (ZonMW) to N.M.v.S. Publisher Copyright: © 2021 The Authors. Chemistry - A European Journal published by Wiley-VCH GmbH
PY - 2021/7/16
Y1 - 2021/7/16
N2 - Wall teichoic acids (WTAs) are important components of the cell wall of the opportunistic Gram-positive bacterium Staphylococcus aureus. WTAs are composed of repeating ribitol phosphate (RboP) residues that are decorated with d-alanine and N-acetyl-d-glucosamine (GlcNAc) modifications, in a seemingly random manner. These WTA-modifications play an important role in shaping the interactions of WTA with the host immune system. Due to the structural heterogeneity of WTAs, it is impossible to isolate pure and well-defined WTA molecules from bacterial sources. Therefore, here synthetic chemistry to assemble a broad library of WTA-fragments, incorporating all possible glycosylation modifications (α-GlcNAc at the RboP C4; β-GlcNAc at the RboP C4; β-GlcNAc at the RboP C3) described for S. aureus WTAs, is reported. DNA-type chemistry, employing ribitol phosphoramidite building blocks, protected with a dimethoxy trityl group, was used to efficiently generate a library of WTA-hexamers. Automated solid phase syntheses were used to assemble a WTA-dodecamer and glycosylated WTA-hexamer. The synthetic fragments have been fully characterized and diagnostic signals were identified to discriminate the different glycosylation patterns. The different glycosylated WTA-fragments were used to probe binding of monoclonal antibodies using WTA-functionalized magnetic beads, revealing the binding specificity of these WTA-specific antibodies and the importance of the specific location of the GlcNAc modifications on the WTA-chains.
AB - Wall teichoic acids (WTAs) are important components of the cell wall of the opportunistic Gram-positive bacterium Staphylococcus aureus. WTAs are composed of repeating ribitol phosphate (RboP) residues that are decorated with d-alanine and N-acetyl-d-glucosamine (GlcNAc) modifications, in a seemingly random manner. These WTA-modifications play an important role in shaping the interactions of WTA with the host immune system. Due to the structural heterogeneity of WTAs, it is impossible to isolate pure and well-defined WTA molecules from bacterial sources. Therefore, here synthetic chemistry to assemble a broad library of WTA-fragments, incorporating all possible glycosylation modifications (α-GlcNAc at the RboP C4; β-GlcNAc at the RboP C4; β-GlcNAc at the RboP C3) described for S. aureus WTAs, is reported. DNA-type chemistry, employing ribitol phosphoramidite building blocks, protected with a dimethoxy trityl group, was used to efficiently generate a library of WTA-hexamers. Automated solid phase syntheses were used to assemble a WTA-dodecamer and glycosylated WTA-hexamer. The synthetic fragments have been fully characterized and diagnostic signals were identified to discriminate the different glycosylation patterns. The different glycosylated WTA-fragments were used to probe binding of monoclonal antibodies using WTA-functionalized magnetic beads, revealing the binding specificity of these WTA-specific antibodies and the importance of the specific location of the GlcNAc modifications on the WTA-chains.
KW - antibodies
KW - automated synthesis
KW - gram-positive bacteria
KW - ribitol phosphate
KW - wall teichoic acids
UR - http://www.scopus.com/inward/record.url?scp=85107542764&partnerID=8YFLogxK
U2 - https://doi.org/10.1002/chem.202101242
DO - https://doi.org/10.1002/chem.202101242
M3 - Article
C2 - 33991006
SN - 0947-6539
VL - 27
SP - 10461
EP - 10469
JO - Chemistry (Weinheim an der Bergstrasse, Germany)
JF - Chemistry (Weinheim an der Bergstrasse, Germany)
IS - 40
ER -