CRC 1340 - Matrix in Vision 

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Deciphering the structure of glycosaminoglycans and their binding properties with cationic metal components of imaging probes

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Project Content

Glycosaminoglycans (GAGs) are a diverse class of polysaccharides that perform a wide variety of functions. Depending on their structure, GAGs bind metal components of cationic imaging probes with varying affinity, making them attractive candidates as targets for in vivo disease imaging. In this project, ion mobility mass spectrometry and gas phase spectroscopy methods will be developed to characterize the structure and cation binding affinity of GAGs. The results obtained will contribute to a better understanding of the role of GAGs in different diseases.

This picture shows the structure of glykosaminoglycans.
© K. Pagel - FU Berlin
Current workflow for depolymerization, separation and characterization of heparin.

Milestones and News

  • The structural modularity of heparan sulfate diastereomers was studied by cryogenic infrared spectroscopy.
  • The binding of gadolinium to heparin oligosaccharides was confirmed by mass spectrometry.
  • Enzymatic depolymerization of large amounts of glycosaminoglycans (GAGs) was optimized.
Exemplary infrared spectrum of a synthetic, sulfated hyaluronic acid derivative, the signals of the diagnostic groups are color-coded
  • Recombinant expression of heparinases is performed in-house.
  • HPLC methods for purification and separation of heparin, hyaluronic acid and chondroitin sulfate were established.
Mass spectrometric detection of transchelation of gadolinium (Gd) in the presence of glycosaminoglycans (GAGs).

Our team

Project Lead:
Prof. Dr. rer. nat. Kevin Pagel

PhD Candidates (PhD) and Postdoctoral Researchers (PostDoc):
Lukasz Polewski (PhD)
Maike Lettow (PhD)
Dr. Michael Götze (PostDoc)
Dr. Andreas Zappe (PostDoc)

left to right: Dr. Michael Götze, MSc Lukasz Polewski, MSc Maike Lettow, Dr. Andreas Zappe

Publications

  • Miller RL, Guimond SE, Schwörer R, Zubkova OV, Tyler PC, Xu Y, Liu J, Chopra P, Boons GJ, Grabarics M, Manz C, Hofmann J, Karlsson NG, Turnbull JE, Struwe WB, Pagel K. Shotgun ion mobility mass spectrometry sequencing of heparan sulfate saccharides. Nat Commun. 2020 Mar 20;11(1):1481. doi: 10.1038/s41467-020-15284-y. PMID: 32198425; PMCID: PMC7083916.
  • Lettow M, Grabarics M, Greis K, Mucha E, Thomas DA, Chopra P, Boons GJ, Karlsson R, Turnbull JE, Meijer G, Miller RL, von Helden G, Pagel K. Cryogenic Infrared Spectroscopy Reveals Structural Modularity in the Vibrational Fingerprints of Heparan Sulfate Diastereomers. Anal Chem. 2020 Aug 4;92(15):10228-10232. doi: 10.1021/acs.analchem.0c02048. Epub 2020 Jul 21. PMID: 32658472.
  • Lettow M, Grabarics M, Mucha E, Thomas DA, Polewski Ł, Freyse J, Rademann J, Meijer G, von Helden G, Pagel K. IR action spectroscopy of glycosaminoglycan oligosaccharides. Anal Bioanal Chem. 2020 Jan;412(3):533-537. doi: 10.1007/s00216-019-02327-7. Epub 2019 Dec 18. PMID: 31853603; PMCID: PMC6992547.
  • Lettow M, Greis K, Grabarics M, Horlebein J, Miller RL, Meijer G, von Helden G, Pagel K. Chondroitin Sulfate Disaccharides in the Gas Phase: Differentiation and Conformational Constraints. J Phys Chem A. 2021 May 27;125(20):4373-4379. doi: 10.1021/acs.jpca.1c02463. Epub 2021 May 12. PMID: 33979516.

Grants and Awards

Berlin University Alliance: "Zwischen den Zellen" (2019)

ERC Consolidator Grant (2019)

CORDIS: H2020 FET Open HS-SEQ. (2020)

Tagesspiegel: Unerforschter Klassiker (2020)

Projects